LOCUS       Exported                8099 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d15 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8138)
  AUTHORS   Stafl K, Travnicek M, Kucerova D, Pecnova L, Krchlikova V, Galikova 
            E, Hejnar J and Trejbalova K
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 5, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8099
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1584
                     /codon_start=1
                     /label=hASCT2-d15
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTGGVGQEVEGMNILGLV
                     VFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEMED
                     VGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTSSS
                     SATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLDFV
                     KIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTVLN
                     VEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPAGD
                     ATVASEKESVM"
     misc_feature    124..150
                     /label=TM1a
                     /note="Garaeva et al., 2018"
     misc_feature    154..222
                     /product="Garaeva et al., 2018"
                     /label=TM1b
     misc_feature    247..351
                     /label=TM2
                     /note="Garaeva et al., 2018"
     misc_feature    376..459
                     /label=TM3
                     /note="Garaeva et al., 2018"
     misc_feature    460..540
                     /label=region N
     misc_feature    541..570
                     /product="Garaeva et al., 2018"
                     /label=TM4a
     misc_feature    583..609
                     /label=TM4b
     misc_feature    619..624
                     /label=d15
     misc_feature    655..705
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    721..837
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    850..948
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    958..1011
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1018..1059
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1075..1116
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1123..1170
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1192..1236
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1261..1299
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1324..1443
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1600..1614
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1615..2331
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2349..2935
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2936..3886
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     LTR             4000..4385
                     /label=dLTR
     V_segment       complement(4386..7129)
                     /label=pLitmus 28i
     promoter        complement(4441..4459)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4441..4459)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4480..4496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4504..4520
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4528..4558)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4655..5243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5354..5863
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5905..6765)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6766..6870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7091..7107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7111..7129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7111..7129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7130..7521
     LTR             7130..7515
                     /label=dLTR
     misc_feature    7522..7539
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7576..7680
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7792..8054
                     /label=p19
                     /note="p19"
     gene            8055..8068
                     /label=fragment p27
                     /note="fragment p27"
ORIGIN
        1 atggtggccg atcctcctcg agactccaag gggctcgcag cggcggagcc caccgccaac
       61 gggggcctgg cgctggcctc catcgaggac caaggcgcgg cagcaggcgg ctactgcggt
      121 tcccgggacc aggtgcgccg ctgccttcga gccaacctgc ttgtgctgct gacagtggtg
      181 gccgtggtgg ccggcgtggc gctgggactg ggggtgtcgg gggccggggg tgcgctggcg
      241 ttgggcccgg agcgcttgag cgccttcgtc ttcccgggcg agctgctgct gcgtctgctg
      301 cggatgatca tcttgccgct ggtggtgtgc agcttgatcg gcggcgccgc cagcctggac
      361 cccggcgcgc tcggccgtct gggcgcctgg gcgctgctct ttttcctggt caccacgctg
      421 ctggcgtcgg cgctcggagt gggcttggcg ctggctctgc agccgggcgc cgcctccgcc
      481 gccatcaacg cctccgtggg agccgcgggc agtgccgaaa atgcccccag caaggaggtg
      541 ctcgattcgt tcctggatct tgcgagaaat atcttccctt ccaacctggt gtcagcagcc
      601 tttcgctcat actctaccgg tggggtgggg caggaggtgg aggggatgaa catcctgggc
      661 ttggtagtgt ttgcaattgt ctttggtgtg gcgctgcgga agctggggcc tgaaggggag
      721 ctgcttatcc gcttcttcaa ctccttcaat gaggccacca tggttctggt ctcctggatc
      781 atgtggtacg cccctgtggg catcatgttc ctggtggctg gcaagatcgt ggagatggag
      841 gatgtgggtt tactctttgc ccgccttggc aagtacattc tgtgctgcct gctgggtcac
      901 gccatccatg ggctcctggt actgcccctc atctacttcc tcttcacccg caaaaacccc
      961 taccgcttcc tgtggggcat cgtgacgccg ctggccactg cctttgggac ctcttccagt
     1021 tccgccacgc tgccgctgat gatgaagtgc gtggaggaga ataatggcgt ggccaagcac
     1081 atcagccgtt tcatcctgcc catcggcgcc accgtcaaca tggacggtgc cgcgctcttc
     1141 cagtgcgtgg ccgcagtgtt cattgcacag ctcagccagc agtccttgga cttcgtaaag
     1201 atcatcacca tcctggtcac ggccacagcg tccagcgtgg gggcagcggg catccctgct
     1261 ggaggtgtcc tcactctggc catcatcctc gaagcagtca acctcccggt cgaccatatc
     1321 tccttgatcc tggctgtgga ctggctagtc gaccggtcct gtaccgtcct caatgtagaa
     1381 ggtgacgctc tgggggcagg actcctccaa aattacgtgg accgtacgga gtcgagaagc
     1441 acagagcctg agttgataca agtgaagagt gagctgcccc tggatccgct gccagtcccc
     1501 actgaggaag gaaaccccct cctcaaacac tatcgggggc ccgcagggga tgccacggtc
     1561 gcctctgaga aggaatcagt catgactagt accggtaccg gaggtggtgg atccgtgagc
     1621 aagggcgccg agctgttcac cggcatcgtg cccatcctga tcgagctgaa tggcgatgtg
     1681 aatggccaca agttcagcgt gagcggcgag ggcgagggcg atgccaccta cggcaagctg
     1741 accctgaagt tcatctgcac caccggcaag ctgcctgtgc cctggcccac cctggtgacc
     1801 accctgagct acggcgtgca gtgcttctca cgctaccccg atcacatgaa gcagcacgac
     1861 ttcttcaaga gcgccatgcc tgagggctac atccaggagc gcaccatctt cttcgaggat
     1921 gacggcaact acaagtcgcg cgccgaggtg aagttcgagg gcgataccct ggtgaatcgc
     1981 atcgagctga ccggcaccga tttcaaggag gatggcaaca tcctgggcaa taagatggag
     2041 tacaactaca acgcccacaa tgtgtacatc atgaccgaca aggccaagaa tggcatcaag
     2101 gtgaacttca agatccgcca caacatcgag gatggcagcg tgcagctggc cgaccactac
     2161 cagcagaata cccccatcgg cgatggccct gtgctgctgc ccgataacca ctacctgtcc
     2221 acccagagcg ccctgtccaa ggaccccaac gagaagcgcg atcacatgat ctacttcggc
     2281 ttcgtgaccg ccgccgccat cacccacggc atggatgagc tgtacaagtg agcggccgcg
     2341 actctagacc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2401 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2461 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2521 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2581 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2641 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2701 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     2761 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     2821 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     2881 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggc
     2941 ggaaggatcc gtcgccaggc agcctgacct cttgacctgc gacgatgagc cgatccatat
     3001 ccccggtgcc atccaaccgc atggactgct gctcgccctc gccgccgaca tgacgatcgt
     3061 tgccggcagc gacaaccttc ccgaactcac cggactggcg atcggcgccc tgatcggccg
     3121 ctctgcggcc gatgtcttcg actcggagac gcacaaccgt ctgacgatcg ccttggccga
     3181 gcccggggcg gccgtcggag caccgatcac tgtcggcttc acgatgcgaa aggacgcagg
     3241 cttcatcggc tcctggcatc gccatgatca gctcatcttc ctcgagctcg agcctcccca
     3301 gcgggacgtc gccgagccgc aggcgttctt ccgccgcacc aacagcgcca tccgccgcct
     3361 gcaggccgcc gaaaccttgg aaagcgcctg cgccgccgcg gcgcaagagg tgcggaagat
     3421 taccggcttc gatcgggtga tgatctatcg cttcgcctcc gacttcagcg gcgaagtgat
     3481 cgcagaggat cggtgcgccg aggtcgagtc aaaactaggc ctgcactatc ctgcctcaac
     3541 cgtgccggcg caggcccgtc ggctctatac catcaacccg gtacggatca ttcccgatat
     3601 caattatcgg ccggtgccgg tcaccccaga cctcaatccg gtcaccgggc ggccgattga
     3661 tcttagcttc gccatcctgc gcagcgtctc gcccgtccat ctggagttca tgcgcaacat
     3721 aggcatgcac ggcacgatgt cgatctcgat tttgcgcggc gagcgactgt ggggattgat
     3781 cgtttgccat caccgaacgc cgtactacgt cgatctcgat ggccgccaag cctgcgagct
     3841 agtcgcccag gttctggcct ggcagatcgg cgtgatggaa gagtgacgcg tatctgaggg
     3901 gactagggtg tgtttaggcg aaaagcgggg cttcggttgt acgcggttag gagtcccctc
     3961 gaggtatggc agatatgctt ttgcataggg agggggaaat gtagtcttaa tcataggtta
     4021 acatgtatat taccaaataa gggaatcgcc tgatgcacca aataaggtat tatatgatcc
     4081 cattggtggc gagggagcga cctgagggca tatgggcgtt aacagaactg tctgtccttg
     4141 cgtcattcct tcctcatcgg atcatgtacg cggcagagta tgattggata acaggatggc
     4201 accattcatc gtggcgcatg ctgattggtg cgactaagga gttatgtaac ccacgaatgt
     4261 acttaagctt gtagttgcta acaataaagg gccattctac ctctcaccac attggtgtgc
     4321 acctgggttg atggccggac cgtcgattcc ctgacgacta cgagcacctg aatgaagcag
     4381 aaggcatcct gcaggaattc ctcgagacgt acggtgcgcg cgatgcattc gaagatctgc
     4441 cctatagtga gtcgtattac gtagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4561 ctggggtgcc taatgagtga gctaactcac attacatgtg agcaaaaggc cagcaaaagg
     4621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4981 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5281 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5341 cctagatcct tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5401 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5461 tcctttctcg ccacgttcgc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5521 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5581 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5641 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     5701 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5761 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaata tttgcttata
     5821 caatcttcct gtttttgggg cttttctgat tatcaaccgg ggtaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggagaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 agttaacaaa aaaaagcccg ccgaagcggg ctttattacc aagcgaagcg ccattcgcca
     6961 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7021 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7081 tcacgacgtt gtaaaacgac ggccagtccg taatacgact cactataggt gtagtcttaa
     7141 tcataggtta acatgtatat taccaaataa gggaatcgcc tgatgcacca aataaggtat
     7201 tatatgatcc cattggtggc gagggagcga cctgagggca tatgggcgtt aacagaactg
     7261 tctgtccttg cgtcattcct tcctcatcgg atcatgtacg cggcagagta tgattggata
     7321 acaggatggc accattcatc gtggcgcatg ctgattggtg cgactaagga gttatgtaac
     7381 ccacgaatgt acttaagctt gtagttgcta acaataaagg gccattctac ctctcaccac
     7441 attggtgtgc acctgggttg atggccggac cgtcgattcc ctgacgacta cgagcacctg
     7501 aatgaagcag aaggcttcat ttggtgaccc cgacgtgatc gttagggaat agtggtcggc
     7561 cacaggcggc gtggcgatcc tgtctccatc cgtctcgtcg accgggaggc gagttcgatg
     7621 accctgatag agggggctgc ggcttaggga ggcagaagct gagcaccgtc ggtaggagct
     7681 ccagggcccg gagcgactga cccctgccga gaactcagag ggtcgtcgga agacggagag
     7741 tgagcccgac gaccacccca ggcacgtctt tggtcggcct gcggatcaag ctagcaagcc
     7801 gtcattaagg tgatttcgtc cgcgtgtaaa acctattgcg ggaaaatctc tccttctaag
     7861 aaggaaatag gggccatgtt gtccctgtta caaaaggaag ggttgcttta gtctccctca
     7921 gatttatatt ctccggggtc ctgggatccc atcactgcgg cgctctccca gcgggcaatg
     7981 gtacttggaa aatcgggaga gttaaaaacc tggggattgg ttttgggggc attgaaggcg
     8041 gctcgagagt aacaggacta acttgaatcg atgcgctacc ggactcagat cttgccacc
//
