LOCUS Exported 8138 bp ds-DNA circular SYN 05-FEB-2021 DEFINITION pFuTraP vector expressing human ASCT2 with N212Q substitution. 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..8138 /organism="recombinant plasmid" /lab_host="Gallus gallus" /mol_type="other DNA" CDS 1..1623 /codon_start=1 /product="ASCT2" /label=hASCT2 /note="human ASCT2" /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYEERQITGTRVKVPV GQEVEGMNILGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGI MFLVAGKIVEMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGI VTPLATAFGTSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAV FIAQLSQQSLDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILA VDWLVDRSCTVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEG NPLLKHYRGPAGDATVASEKESVM" 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 mat_peptide 610..681 /label=region C misc_feature 694..744 /label=TM4c /note="Garaeva et al., 2018" misc_feature 760..876 /label=TM5 /note="Garaeva et al., 2018" misc_feature 889..987 /label=TM6 /note="Garaeva et al., 2018" misc_feature 997..1050 /label=HP1a /note="Garaeva et al., 2018" misc_feature 1057..1098 /label=HP1b /note="Garaeva et al., 2018" misc_feature 1114..1155 /label=TM7a /note="Garaeva et al., 2018" misc_feature 1162..1209 /label=TM7b /note="Garaeva et al., 2018" misc_feature 1231..1275 /label=HP2a /note="Garaeva et al., 2018" misc_feature 1300..1338 /label=HP2b /note="Garaeva et al., 2018" misc_feature 1363..1482 /label=TM8 /note="Garaeva et al., 2018" CDS 1639..1653 /codon_start=1 /label=GGGGS linker /translation="GGGGS" CDS 1654..2370 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG FVTAAAITHGMDELYK" misc_feature 2388..2974 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 2975..3925 /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 4039..4424 /label=dLTR V_segment complement(4425..7168) /label=pLitmus 28i promoter complement(4480..4498) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4519..4535) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4543..4559 /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(4567..4597) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4612..4633 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4694..5282) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" rep_origin 5393..5902 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(5944..6804) /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(6805..6909) /gene="bla" /label=AmpR promoter primer_bind 7130..7146 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7150..7168 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" LTR 7169..7560 misc_feature 7561..7578 /label=PBS /note="the same as for RCAS" misc_feature 7615..7719 /label=packaging signal /note="according to Banks JD and Linial ML, J. Virol., 2000" gene 7831..8093 /label=p19 /note="p19" gene 8094..8107 /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 aggcagatca ccggaaccag ggtgaaggtg 661 cccgtggggc aggaggtgga ggggatgaac atcctgggct tggtagtgtt tgcaattgtc 721 tttggtgtgg cgctgcggaa gctggggcct gaaggggagc tgcttatccg cttcttcaac 781 tccttcaatg aggccaccat ggttctggtc tcctggatca tgtggtacgc ccctgtgggc 841 atcatgttcc tggtggctgg caagatcgtg gagatggagg atgtgggttt actctttgcc 901 cgccttggca agtacattct gtgctgcctg ctgggtcacg ccatccatgg gctcctggta 961 ctgcccctca tctacttcct cttcacccgc aaaaacccct accgcttcct gtggggcatc 1021 gtgacgccgc tggccactgc ctttgggacc tcttccagtt ccgccacgct gccgctgatg 1081 atgaagtgcg tggaggagaa taatggcgtg gccaagcaca tcagccgttt catcctgccc 1141 atcggcgcca ccgtcaacat ggacggtgcc gcgctcttcc agtgcgtggc cgcagtgttc 1201 attgcacagc tcagccagca gtccttggac ttcgtaaaga tcatcaccat cctggtcacg 1261 gccacagcgt ccagcgtggg ggcagcgggc atccctgctg gaggtgtcct cactctggcc 1321 atcatcctcg aagcagtcaa cctcccggtc gaccatatct ccttgatcct ggctgtggac 1381 tggctagtcg accggtcctg taccgtcctc aatgtagaag gtgacgctct gggggcagga 1441 ctcctccaaa attacgtgga ccgtacggag tcgagaagca cagagcctga gttgatacaa 1501 gtgaagagtg agctgcccct ggatccgctg ccagtcccca ctgaggaagg aaaccccctc 1561 ctcaaacact atcgggggcc cgcaggggat gccacggtcg cctctgagaa ggaatcagtc 1621 atgactagta ccggtaccgg aggtggtgga tccgtgagca agggcgccga gctgttcacc 1681 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg 1741 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 1801 accggcaagc tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag 1861 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct 1921 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc 1981 gccgaggtga agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat 2041 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat 2101 gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac 2161 aacatcgagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc 2221 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag 2281 gaccccaacg agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc 2341 acccacggca tggatgagct gtacaagtga gcggccgcga ctctagaccc ctctccctcc 2401 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat 2461 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct 2521 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg 2581 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta 2641 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag 2701 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg 2761 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat 2821 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca 2881 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc 2941 ctttgaaaaa cacgatgata atatggccac aaccatggcg gaaggatccg tcgccaggca 3001 gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca 3061 tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc 3121 cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga 3181 ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc 3241 accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg 3301 ccatgatcag ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca 3361 ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga 3421 aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat 3481 gatctatcgc ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga 3541 ggtcgagtca aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc aggcccgtcg 3601 gctctatacc atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt 3661 caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg 3721 cagcgtctcg cccgtccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc 3781 gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc 3841 gtactacgtc gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg ttctggcctg 3901 gcagatcggc gtgatggaag agtgacgcgt atctgagggg actagggtgt gtttaggcga 3961 aaagcggggc ttcggttgta cgcggttagg agtcccctcg aggtatggca gatatgcttt 4021 tgcataggga gggggaaatg tagtcttaat cataggttaa catgtatatt accaaataag 4081 ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg agggagcgac 4141 ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt cctcatcgga 4201 tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg tggcgcatgc 4261 tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg tagttgctaa 4321 caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga tggccggacc 4381 gtcgattccc tgacgactac gagcacctga atgaagcaga aggcatcctg caggaattcc 4441 tcgagacgta cggtgcgcgc gatgcattcg aagatctgcc ctatagtgag tcgtattacg 4501 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4621 ctaactcaca ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttacgcgccc 5401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 5461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgct 5521 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 5581 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat 5641 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 5701 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc 5761 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 5821 acaaaatatt aacgtttaca atttaaatat ttgcttatac aatcttcctg tttttggggc 5881 ttttctgatt atcaaccggg gtaaatcaat ctaaagtata tatgagtaaa cttggtctga 5941 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6001 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6061 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 6121 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 6181 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 6241 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 6301 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 6361 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 6421 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 6481 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 6541 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt 6601 gctcatcatt ggagaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 6661 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 6721 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 6781 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 6841 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 6901 ggttccgcgc acatttcccc gaaaagtgcc acctgacgta gttaacaaaa aaaagcccgc 6961 cgaagcgggc tttattacca agcgaagcgc cattcgccat tcaggctgcg caactgttgg 7021 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct 7081 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 7141 gccagtccgt aatacgactc actataggtg tagtcttaat cataggttaa catgtatatt 7201 accaaataag ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg 7261 agggagcgac ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt 7321 cctcatcgga tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg 7381 tggcgcatgc tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg 7441 tagttgctaa caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga 7501 tggccggacc gtcgattccc tgacgactac gagcacctga atgaagcaga aggcttcatt 7561 tggtgacccc gacgtgatcg ttagggaata gtggtcggcc acaggcggcg tggcgatcct 7621 gtctccatcc gtctcgtcga ccgggaggcg agttcgatga ccctgataga gggggctgcg 7681 gcttagggag gcagaagctg agcaccgtcg gtaggagctc cagggcccgg agcgactgac 7741 ccctgccgag aactcagagg gtcgtcggaa gacggagagt gagcccgacg accaccccag 7801 gcacgtcttt ggtcggcctg cggatcaagc tagcaagccg tcattaaggt gatttcgtcc 7861 gcgtgtaaaa cctattgcgg gaaaatctct ccttctaaga aggaaatagg ggccatgttg 7921 tccctgttac aaaaggaagg gttgctttag tctccctcag atttatattc tccggggtcc 7981 tgggatccca tcactgcggc gctctcccag cgggcaatgg tacttggaaa atcgggagag 8041 ttaaaaacct ggggattggt tttgggggca ttgaaggcgg ctcgagagta acaggactaa 8101 cttgaatcga tgcgctaccg gactcagatc ttgccacc //