LOCUS Exported 8123 bp ds-DNA circular SYN 05-FEB-2021 DEFINITION pFuTraP vector expressing human ASCT2 with d7 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..8123 /organism="recombinant plasmid" /lab_host="Gallus gallus" /mol_type="other DNA" CDS 1..1608 /codon_start=1 /label=hASCT2-d7 /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYEEGGVKVPVGQEVE GMNILGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVA GKIVEMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLA TAFGTSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQL SQQSLDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLV DRSCTVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLK HYRGPAGDATVASEKESVM" 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 631..636 /label=d7 misc_feature 679..729 /label=TM4c /note="Garaeva et al., 2018" misc_feature 745..861 /label=TM5 /note="Garaeva et al., 2018" misc_feature 874..972 /label=TM6 /note="Garaeva et al., 2018" misc_feature 982..1035 /label=HP1a /note="Garaeva et al., 2018" misc_feature 1042..1083 /label=HP1b /note="Garaeva et al., 2018" misc_feature 1099..1140 /label=TM7a /note="Garaeva et al., 2018" misc_feature 1147..1194 /label=TM7b /note="Garaeva et al., 2018" misc_feature 1216..1260 /label=HP2a /note="Garaeva et al., 2018" misc_feature 1285..1323 /label=HP2b /note="Garaeva et al., 2018" misc_feature 1348..1467 /label=TM8 /note="Garaeva et al., 2018" CDS 1624..1638 /codon_start=1 /label=GGGGS linker /translation="GGGGS" CDS 1639..2355 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG FVTAAAITHGMDELYK" misc_feature 2373..2959 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 2960..3910 /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 4024..4409 /label=dLTR V_segment complement(4410..7153) /label=pLitmus 28i promoter complement(4465..4483) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4504..4520) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4528..4544 /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(4552..4582) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4597..4618 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4679..5267) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" rep_origin 5378..5887 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(5929..6789) /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(6790..6894) /gene="bla" /label=AmpR promoter primer_bind 7115..7131 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7135..7153 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" LTR 7154..7545 misc_feature 7546..7563 /label=PBS /note="the same as for RCAS" misc_feature 7600..7704 /label=packaging signal /note="according to Banks JD and Linial ML, J. Virol., 2000" gene 7816..8078 /label=p19 /note="p19" gene 8079..8092 /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 ggaggagtta aggtgcccgt ggggcaggag 661 gtggagggga tgaacatcct gggcttggta gtgtttgcaa ttgtctttgg tgtggcgctg 721 cggaagctgg ggcctgaagg ggagctgctt atccgcttct tcaactcctt caatgaggcc 781 accatggttc tggtctcctg gatcatgtgg tacgcccctg tgggcatcat gttcctggtg 841 gctggcaaga tcgtggagat ggaggatgtg ggtttactct ttgcccgcct tggcaagtac 901 attctgtgct gcctgctggg tcacgccatc catgggctcc tggtactgcc cctcatctac 961 ttcctcttca cccgcaaaaa cccctaccgc ttcctgtggg gcatcgtgac gccgctggcc 1021 actgcctttg ggacctcttc cagttccgcc acgctgccgc tgatgatgaa gtgcgtggag 1081 gagaataatg gcgtggccaa gcacatcagc cgtttcatcc tgcccatcgg cgccaccgtc 1141 aacatggacg gtgccgcgct cttccagtgc gtggccgcag tgttcattgc acagctcagc 1201 cagcagtcct tggacttcgt aaagatcatc accatcctgg tcacggccac agcgtccagc 1261 gtgggggcag cgggcatccc tgctggaggt gtcctcactc tggccatcat cctcgaagca 1321 gtcaacctcc cggtcgacca tatctccttg atcctggctg tggactggct agtcgaccgg 1381 tcctgtaccg tcctcaatgt agaaggtgac gctctggggg caggactcct ccaaaattac 1441 gtggaccgta cggagtcgag aagcacagag cctgagttga tacaagtgaa gagtgagctg 1501 cccctggatc cgctgccagt ccccactgag gaaggaaacc ccctcctcaa acactatcgg 1561 gggcccgcag gggatgccac ggtcgcctct gagaaggaat cagtcatgac tagtaccggt 1621 accggaggtg gtggatccgt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc 1681 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag 1741 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct 1801 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac 1861 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag 1921 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc 1981 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc 2041 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc 2101 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc 2161 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg 2221 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag 2281 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat 2341 gagctgtaca agtgagcggc cgcgactcta gacccctctc cctccccccc ccctaacgtt 2401 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 2461 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 2521 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 2581 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 2641 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 2701 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga 2761 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 2821 cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg 2881 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga 2941 tgataatatg gccacaacca tggcggaagg atccgtcgcc aggcagcctg acctcttgac 3001 ctgcgacgat gagccgatcc atatccccgg tgccatccaa ccgcatggac tgctgctcgc 3061 cctcgccgcc gacatgacga tcgttgccgg cagcgacaac cttcccgaac tcaccggact 3121 ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc ttcgactcgg agacgcacaa 3181 ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc ggagcaccga tcactgtcgg 3241 cttcacgatg cgaaaggacg caggcttcat cggctcctgg catcgccatg atcagctcat 3301 cttcctcgag ctcgagcctc cccagcggga cgtcgccgag ccgcaggcgt tcttccgccg 3361 caccaacagc gccatccgcc gcctgcaggc cgccgaaacc ttggaaagcg cctgcgccgc 3421 cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg gtgatgatct atcgcttcgc 3481 ctccgacttc agcggcgaag tgatcgcaga ggatcggtgc gccgaggtcg agtcaaaact 3541 aggcctgcac tatcctgcct caaccgtgcc ggcgcaggcc cgtcggctct ataccatcaa 3601 cccggtacgg atcattcccg atatcaatta tcggccggtg ccggtcaccc cagacctcaa 3661 tccggtcacc gggcggccga ttgatcttag cttcgccatc ctgcgcagcg tctcgcccgt 3721 ccatctggag ttcatgcgca acataggcat gcacggcacg atgtcgatct cgattttgcg 3781 cggcgagcga ctgtggggat tgatcgtttg ccatcaccga acgccgtact acgtcgatct 3841 cgatggccgc caagcctgcg agctagtcgc ccaggttctg gcctggcaga tcggcgtgat 3901 ggaagagtga cgcgtatctg aggggactag ggtgtgttta ggcgaaaagc ggggcttcgg 3961 ttgtacgcgg ttaggagtcc cctcgaggta tggcagatat gcttttgcat agggaggggg 4021 aaatgtagtc ttaatcatag gttaacatgt atattaccaa ataagggaat cgcctgatgc 4081 accaaataag gtattatatg atcccattgg tggcgaggga gcgacctgag ggcatatggg 4141 cgttaacaga actgtctgtc cttgcgtcat tccttcctca tcggatcatg tacgcggcag 4201 agtatgattg gataacagga tggcaccatt catcgtggcg catgctgatt ggtgcgacta 4261 aggagttatg taacccacga atgtacttaa gcttgtagtt gctaacaata aagggccatt 4321 ctacctctca ccacattggt gtgcacctgg gttgatggcc ggaccgtcga ttccctgacg 4381 actacgagca cctgaatgaa gcagaaggca tcctgcagga attcctcgag acgtacggtg 4441 cgcgcgatgc attcgaagat ctgccctata gtgagtcgta ttacgtagct tggcgtaatc 4501 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4561 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaca 4621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5101 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 5161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 5221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5341 tgagattatc aaaaaggatc ttcacctaga tccttttacg cgccctgtag cggcgcatta 5401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 5461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgctttccc cgtcaagctc 5521 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 5581 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 5641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 5701 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt 5761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt 5821 ttacaattta aatatttgct tatacaatct tcctgttttt ggggcttttc tgattatcaa 5881 ccggggtaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6541 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaga 6601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6901 tccccgaaaa gtgccacctg acgtagttaa caaaaaaaag cccgccgaag cgggctttat 6961 taccaagcga agcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 7021 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 7081 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tccgtaatac 7141 gactcactat aggtgtagtc ttaatcatag gttaacatgt atattaccaa ataagggaat 7201 cgcctgatgc accaaataag gtattatatg atcccattgg tggcgaggga gcgacctgag 7261 ggcatatggg cgttaacaga actgtctgtc cttgcgtcat tccttcctca tcggatcatg 7321 tacgcggcag agtatgattg gataacagga tggcaccatt catcgtggcg catgctgatt 7381 ggtgcgacta aggagttatg taacccacga atgtacttaa gcttgtagtt gctaacaata 7441 aagggccatt ctacctctca ccacattggt gtgcacctgg gttgatggcc ggaccgtcga 7501 ttccctgacg actacgagca cctgaatgaa gcagaaggct tcatttggtg accccgacgt 7561 gatcgttagg gaatagtggt cggccacagg cggcgtggcg atcctgtctc catccgtctc 7621 gtcgaccggg aggcgagttc gatgaccctg atagaggggg ctgcggctta gggaggcaga 7681 agctgagcac cgtcggtagg agctccaggg cccggagcga ctgacccctg ccgagaactc 7741 agagggtcgt cggaagacgg agagtgagcc cgacgaccac cccaggcacg tctttggtcg 7801 gcctgcggat caagctagca agccgtcatt aaggtgattt cgtccgcgtg taaaacctat 7861 tgcgggaaaa tctctccttc taagaaggaa ataggggcca tgttgtccct gttacaaaag 7921 gaagggttgc tttagtctcc ctcagattta tattctccgg ggtcctggga tcccatcact 7981 gcggcgctct cccagcgggc aatggtactt ggaaaatcgg gagagttaaa aacctgggga 8041 ttggttttgg gggcattgaa ggcggctcga gagtaacagg actaacttga atcgatgcgc 8101 taccggactc agatcttgcc acc //