LOCUS       Exported                8129 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d5 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..8129
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1614
                     /codon_start=1
                     /label=hASCT2-d5
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYEERGGRVKVPVGQE
                     VEGMNILGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFL
                     VAGKIVEMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTP
                     LATAFGTSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIA
                     QLSQQSLDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDW
                     LVDRSCTVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPL
                     LKHYRGPAGDATVASEKESVM"
     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    634..639
                     /label=d5
     misc_feature    685..735
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    751..867
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    880..978
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    988..1041
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1048..1089
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1105..1146
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1153..1200
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1222..1266
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1291..1329
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1354..1473
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1630..1644
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1645..2361
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2379..2965
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2966..3916
                     /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             4030..4415
                     /label=dLTR
     V_segment       complement(4416..7159)
                     /label=pLitmus 28i
     promoter        complement(4471..4489)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4510..4526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4534..4550
                     /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(4558..4588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4603..4624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4685..5273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5384..5893
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5935..6795)
                     /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(6796..6900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7121..7137
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7141..7159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7160..7551
     misc_feature    7552..7569
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7606..7710
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7822..8084
                     /label=p19
                     /note="p19"
     gene            8085..8098
                     /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 actctaccac ctatgaagag aggggaggac gcgtgaaggt gcccgtgggg
      661 caggaggtgg aggggatgaa catcctgggc ttggtagtgt ttgcaattgt ctttggtgtg
      721 gcgctgcgga agctggggcc tgaaggggag ctgcttatcc gcttcttcaa ctccttcaat
      781 gaggccacca tggttctggt ctcctggatc atgtggtacg cccctgtggg catcatgttc
      841 ctggtggctg gcaagatcgt ggagatggag gatgtgggtt tactctttgc ccgccttggc
      901 aagtacattc tgtgctgcct gctgggtcac gccatccatg ggctcctggt actgcccctc
      961 atctacttcc tcttcacccg caaaaacccc taccgcttcc tgtggggcat cgtgacgccg
     1021 ctggccactg cctttgggac ctcttccagt tccgccacgc tgccgctgat gatgaagtgc
     1081 gtggaggaga ataatggcgt ggccaagcac atcagccgtt tcatcctgcc catcggcgcc
     1141 accgtcaaca tggacggtgc cgcgctcttc cagtgcgtgg ccgcagtgtt cattgcacag
     1201 ctcagccagc agtccttgga cttcgtaaag atcatcacca tcctggtcac ggccacagcg
     1261 tccagcgtgg gggcagcggg catccctgct ggaggtgtcc tcactctggc catcatcctc
     1321 gaagcagtca acctcccggt cgaccatatc tccttgatcc tggctgtgga ctggctagtc
     1381 gaccggtcct gtaccgtcct caatgtagaa ggtgacgctc tgggggcagg actcctccaa
     1441 aattacgtgg accgtacgga gtcgagaagc acagagcctg agttgataca agtgaagagt
     1501 gagctgcccc tggatccgct gccagtcccc actgaggaag gaaaccccct cctcaaacac
     1561 tatcgggggc ccgcagggga tgccacggtc gcctctgaga aggaatcagt catgactagt
     1621 accggtaccg gaggtggtgg atccgtgagc aagggcgccg agctgttcac cggcatcgtg
     1681 cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt gagcggcgag
     1741 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1801 ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca gtgcttctca
     1861 cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc tgagggctac
     1921 atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg cgccgaggtg
     1981 aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga tttcaaggag
     2041 gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa tgtgtacatc
     2101 atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca caacatcgag
     2161 gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg cgatggccct
     2221 gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa ggaccccaac
     2281 gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat cacccacggc
     2341 atggatgagc tgtacaagtg agcggccgcg actctagacc cctctccctc ccccccccct
     2401 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2461 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2521 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     2581 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     2641 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     2701 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     2761 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     2821 gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag
     2881 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     2941 acacgatgat aatatggcca caaccatggc ggaaggatcc gtcgccaggc agcctgacct
     3001 cttgacctgc gacgatgagc cgatccatat ccccggtgcc atccaaccgc atggactgct
     3061 gctcgccctc gccgccgaca tgacgatcgt tgccggcagc gacaaccttc ccgaactcac
     3121 cggactggcg atcggcgccc tgatcggccg ctctgcggcc gatgtcttcg actcggagac
     3181 gcacaaccgt ctgacgatcg ccttggccga gcccggggcg gccgtcggag caccgatcac
     3241 tgtcggcttc acgatgcgaa aggacgcagg cttcatcggc tcctggcatc gccatgatca
     3301 gctcatcttc ctcgagctcg agcctcccca gcgggacgtc gccgagccgc aggcgttctt
     3361 ccgccgcacc aacagcgcca tccgccgcct gcaggccgcc gaaaccttgg aaagcgcctg
     3421 cgccgccgcg gcgcaagagg tgcggaagat taccggcttc gatcgggtga tgatctatcg
     3481 cttcgcctcc gacttcagcg gcgaagtgat cgcagaggat cggtgcgccg aggtcgagtc
     3541 aaaactaggc ctgcactatc ctgcctcaac cgtgccggcg caggcccgtc ggctctatac
     3601 catcaacccg gtacggatca ttcccgatat caattatcgg ccggtgccgg tcaccccaga
     3661 cctcaatccg gtcaccgggc ggccgattga tcttagcttc gccatcctgc gcagcgtctc
     3721 gcccgtccat ctggagttca tgcgcaacat aggcatgcac ggcacgatgt cgatctcgat
     3781 tttgcgcggc gagcgactgt ggggattgat cgtttgccat caccgaacgc cgtactacgt
     3841 cgatctcgat ggccgccaag cctgcgagct agtcgcccag gttctggcct ggcagatcgg
     3901 cgtgatggaa gagtgacgcg tatctgaggg gactagggtg tgtttaggcg aaaagcgggg
     3961 cttcggttgt acgcggttag gagtcccctc gaggtatggc agatatgctt ttgcataggg
     4021 agggggaaat gtagtcttaa tcataggtta acatgtatat taccaaataa gggaatcgcc
     4081 tgatgcacca aataaggtat tatatgatcc cattggtggc gagggagcga cctgagggca
     4141 tatgggcgtt aacagaactg tctgtccttg cgtcattcct tcctcatcgg atcatgtacg
     4201 cggcagagta tgattggata acaggatggc accattcatc gtggcgcatg ctgattggtg
     4261 cgactaagga gttatgtaac ccacgaatgt acttaagctt gtagttgcta acaataaagg
     4321 gccattctac ctctcaccac attggtgtgc acctgggttg atggccggac cgtcgattcc
     4381 ctgacgacta cgagcacctg aatgaagcag aaggcatcct gcaggaattc ctcgagacgt
     4441 acggtgcgcg cgatgcattc gaagatctgc cctatagtga gtcgtattac gtagcttggc
     4501 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4561 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4621 attacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttacgcgcc ctgtagcggc
     5401 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5461 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc tttccccgtc
     5521 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5581 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5641 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5701 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5761 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5821 taacgtttac aatttaaata tttgcttata caatcttcct gtttttgggg cttttctgat
     5881 tatcaaccgg ggtaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggagaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt agttaacaaa aaaaagcccg ccgaagcggg
     6961 ctttattacc aagcgaagcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7021 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     7081 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtccg
     7141 taatacgact cactataggt gtagtcttaa tcataggtta acatgtatat taccaaataa
     7201 gggaatcgcc tgatgcacca aataaggtat tatatgatcc cattggtggc gagggagcga
     7261 cctgagggca tatgggcgtt aacagaactg tctgtccttg cgtcattcct tcctcatcgg
     7321 atcatgtacg cggcagagta tgattggata acaggatggc accattcatc gtggcgcatg
     7381 ctgattggtg cgactaagga gttatgtaac ccacgaatgt acttaagctt gtagttgcta
     7441 acaataaagg gccattctac ctctcaccac attggtgtgc acctgggttg atggccggac
     7501 cgtcgattcc ctgacgacta cgagcacctg aatgaagcag aaggcttcat ttggtgaccc
     7561 cgacgtgatc gttagggaat agtggtcggc cacaggcggc gtggcgatcc tgtctccatc
     7621 cgtctcgtcg accgggaggc gagttcgatg accctgatag agggggctgc ggcttaggga
     7681 ggcagaagct gagcaccgtc ggtaggagct ccagggcccg gagcgactga cccctgccga
     7741 gaactcagag ggtcgtcgga agacggagag tgagcccgac gaccacccca ggcacgtctt
     7801 tggtcggcct gcggatcaag ctagcaagcc gtcattaagg tgatttcgtc cgcgtgtaaa
     7861 acctattgcg ggaaaatctc tccttctaag aaggaaatag gggccatgtt gtccctgtta
     7921 caaaaggaag ggttgcttta gtctccctca gatttatatt ctccggggtc ctgggatccc
     7981 atcactgcgg cgctctccca gcgggcaatg gtacttggaa aatcgggaga gttaaaaacc
     8041 tggggattgg ttttgggggc attgaaggcg gctcgagagt aacaggacta acttgaatcg
     8101 atgcgctacc ggactcagat cttgccacc
//
