LOCUS       Exported                8087 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d19 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..8087
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1572
                     /codon_start=1
                     /label=hASCT2-d19
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYGGQEVEGMNILGLVVFAI
                     VFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEMEDVGLL
                     FARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTSSSSATL
                     PLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLDFVKIIT
                     ILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTVLNVEGD
                     ALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPAGDATVA
                     SEKESVM"
     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    613..618
                     /label=d19
     misc_feature    643..693
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    709..825
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    838..936
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    946..999
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1006..1047
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1063..1104
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1111..1158
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1180..1224
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1249..1287
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1312..1431
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1588..1602
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1603..2319
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2337..2923
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2924..3874
                     /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             3988..4373
                     /label=dLTR
     V_segment       complement(4374..7117)
                     /label=pLitmus 28i
     promoter        complement(4429..4447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4429..4447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4468..4484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4492..4508
                     /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(4516..4546)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4561..4582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5342..5851
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5893..6753)
                     /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(6754..6858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7079..7095
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7099..7117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7099..7117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7118..7509
     LTR             7118..7503
                     /label=dLTR
     misc_feature    7510..7527
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7564..7668
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7780..8042
                     /label=p19
                     /note="p19"
     gene            8043..8056
                     /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 acggtggcca agaggtggag gggatgaaca tcctgggctt ggtagtgttt
      661 gcaattgtct ttggtgtggc gctgcggaag ctggggcctg aaggggagct gcttatccgc
      721 ttcttcaact ccttcaatga ggccaccatg gttctggtct cctggatcat gtggtacgcc
      781 cctgtgggca tcatgttcct ggtggctggc aagatcgtgg agatggagga tgtgggttta
      841 ctctttgccc gccttggcaa gtacattctg tgctgcctgc tgggtcacgc catccatggg
      901 ctcctggtac tgcccctcat ctacttcctc ttcacccgca aaaaccccta ccgcttcctg
      961 tggggcatcg tgacgccgct ggccactgcc tttgggacct cttccagttc cgccacgctg
     1021 ccgctgatga tgaagtgcgt ggaggagaat aatggcgtgg ccaagcacat cagccgtttc
     1081 atcctgccca tcggcgccac cgtcaacatg gacggtgccg cgctcttcca gtgcgtggcc
     1141 gcagtgttca ttgcacagct cagccagcag tccttggact tcgtaaagat catcaccatc
     1201 ctggtcacgg ccacagcgtc cagcgtgggg gcagcgggca tccctgctgg aggtgtcctc
     1261 actctggcca tcatcctcga agcagtcaac ctcccggtcg accatatctc cttgatcctg
     1321 gctgtggact ggctagtcga ccggtcctgt accgtcctca atgtagaagg tgacgctctg
     1381 ggggcaggac tcctccaaaa ttacgtggac cgtacggagt cgagaagcac agagcctgag
     1441 ttgatacaag tgaagagtga gctgcccctg gatccgctgc cagtccccac tgaggaagga
     1501 aaccccctcc tcaaacacta tcgggggccc gcaggggatg ccacggtcgc ctctgagaag
     1561 gaatcagtca tgactagtac cggtaccgga ggtggtggat ccgtgagcaa gggcgccgag
     1621 ctgttcaccg gcatcgtgcc catcctgatc gagctgaatg gcgatgtgaa tggccacaag
     1681 ttcagcgtga gcggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1741 atctgcacca ccggcaagct gcctgtgccc tggcccaccc tggtgaccac cctgagctac
     1801 ggcgtgcagt gcttctcacg ctaccccgat cacatgaagc agcacgactt cttcaagagc
     1861 gccatgcctg agggctacat ccaggagcgc accatcttct tcgaggatga cggcaactac
     1921 aagtcgcgcg ccgaggtgaa gttcgagggc gataccctgg tgaatcgcat cgagctgacc
     1981 ggcaccgatt tcaaggagga tggcaacatc ctgggcaata agatggagta caactacaac
     2041 gcccacaatg tgtacatcat gaccgacaag gccaagaatg gcatcaaggt gaacttcaag
     2101 atccgccaca acatcgagga tggcagcgtg cagctggccg accactacca gcagaatacc
     2161 cccatcggcg atggccctgt gctgctgccc gataaccact acctgtccac ccagagcgcc
     2221 ctgtccaagg accccaacga gaagcgcgat cacatgatct acttcggctt cgtgaccgcc
     2281 gccgccatca cccacggcat ggatgagctg tacaagtgag cggccgcgac tctagacccc
     2341 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2401 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2461 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2521 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2581 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2641 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2701 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2761 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
     2821 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2881 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggcgg aaggatccgt
     2941 cgccaggcag cctgacctct tgacctgcga cgatgagccg atccatatcc ccggtgccat
     3001 ccaaccgcat ggactgctgc tcgccctcgc cgccgacatg acgatcgttg ccggcagcga
     3061 caaccttccc gaactcaccg gactggcgat cggcgccctg atcggccgct ctgcggccga
     3121 tgtcttcgac tcggagacgc acaaccgtct gacgatcgcc ttggccgagc ccggggcggc
     3181 cgtcggagca ccgatcactg tcggcttcac gatgcgaaag gacgcaggct tcatcggctc
     3241 ctggcatcgc catgatcagc tcatcttcct cgagctcgag cctccccagc gggacgtcgc
     3301 cgagccgcag gcgttcttcc gccgcaccaa cagcgccatc cgccgcctgc aggccgccga
     3361 aaccttggaa agcgcctgcg ccgccgcggc gcaagaggtg cggaagatta ccggcttcga
     3421 tcgggtgatg atctatcgct tcgcctccga cttcagcggc gaagtgatcg cagaggatcg
     3481 gtgcgccgag gtcgagtcaa aactaggcct gcactatcct gcctcaaccg tgccggcgca
     3541 ggcccgtcgg ctctatacca tcaacccggt acggatcatt cccgatatca attatcggcc
     3601 ggtgccggtc accccagacc tcaatccggt caccgggcgg ccgattgatc ttagcttcgc
     3661 catcctgcgc agcgtctcgc ccgtccatct ggagttcatg cgcaacatag gcatgcacgg
     3721 cacgatgtcg atctcgattt tgcgcggcga gcgactgtgg ggattgatcg tttgccatca
     3781 ccgaacgccg tactacgtcg atctcgatgg ccgccaagcc tgcgagctag tcgcccaggt
     3841 tctggcctgg cagatcggcg tgatggaaga gtgacgcgta tctgagggga ctagggtgtg
     3901 tttaggcgaa aagcggggct tcggttgtac gcggttagga gtcccctcga ggtatggcag
     3961 atatgctttt gcatagggag ggggaaatgt agtcttaatc ataggttaac atgtatatta
     4021 ccaaataagg gaatcgcctg atgcaccaaa taaggtatta tatgatccca ttggtggcga
     4081 gggagcgacc tgagggcata tgggcgttaa cagaactgtc tgtccttgcg tcattccttc
     4141 ctcatcggat catgtacgcg gcagagtatg attggataac aggatggcac cattcatcgt
     4201 ggcgcatgct gattggtgcg actaaggagt tatgtaaccc acgaatgtac ttaagcttgt
     4261 agttgctaac aataaagggc cattctacct ctcaccacat tggtgtgcac ctgggttgat
     4321 ggccggaccg tcgattccct gacgactacg agcacctgaa tgaagcagaa ggcatcctgc
     4381 aggaattcct cgagacgtac ggtgcgcgcg atgcattcga agatctgccc tatagtgagt
     4441 cgtattacgt agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4561 atgagtgagc taactcacat tacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5461 acgttcgctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5521 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     5581 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5641 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     5701 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5761 cgaattttaa caaaatatta acgtttacaa tttaaatatt tgcttataca atcttcctgt
     5821 ttttggggct tttctgatta tcaaccgggg taaatcaatc taaagtatat atgagtaaac
     5881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6481 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     6541 tttaaaagtg ctcatcattg gagaacgttc ttcggggcga aaactctcaa ggatcttacc
     6601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6841 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtag ttaacaaaaa
     6901 aaagcccgcc gaagcgggct ttattaccaa gcgaagcgcc attcgccatt caggctgcgc
     6961 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     7021 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     7081 aaaacgacgg ccagtccgta atacgactca ctataggtgt agtcttaatc ataggttaac
     7141 atgtatatta ccaaataagg gaatcgcctg atgcaccaaa taaggtatta tatgatccca
     7201 ttggtggcga gggagcgacc tgagggcata tgggcgttaa cagaactgtc tgtccttgcg
     7261 tcattccttc ctcatcggat catgtacgcg gcagagtatg attggataac aggatggcac
     7321 cattcatcgt ggcgcatgct gattggtgcg actaaggagt tatgtaaccc acgaatgtac
     7381 ttaagcttgt agttgctaac aataaagggc cattctacct ctcaccacat tggtgtgcac
     7441 ctgggttgat ggccggaccg tcgattccct gacgactacg agcacctgaa tgaagcagaa
     7501 ggcttcattt ggtgaccccg acgtgatcgt tagggaatag tggtcggcca caggcggcgt
     7561 ggcgatcctg tctccatccg tctcgtcgac cgggaggcga gttcgatgac cctgatagag
     7621 ggggctgcgg cttagggagg cagaagctga gcaccgtcgg taggagctcc agggcccgga
     7681 gcgactgacc cctgccgaga actcagaggg tcgtcggaag acggagagtg agcccgacga
     7741 ccaccccagg cacgtctttg gtcggcctgc ggatcaagct agcaagccgt cattaaggtg
     7801 atttcgtccg cgtgtaaaac ctattgcggg aaaatctctc cttctaagaa ggaaataggg
     7861 gccatgttgt ccctgttaca aaaggaaggg ttgctttagt ctccctcaga tttatattct
     7921 ccggggtcct gggatcccat cactgcggcg ctctcccagc gggcaatggt acttggaaaa
     7981 tcgggagagt taaaaacctg gggattggtt ttgggggcat tgaaggcggc tcgagagtaa
     8041 caggactaac ttgaatcgat gcgctaccgg actcagatct tgccacc
//
