LOCUS Exported 8078 bp ds-DNA circular SYN 05-FEB-2021 DEFINITION pFuTraP vector expressing human ASCT2 with d22 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..8078 /organism="recombinant plasmid" /lab_host="Gallus gallus" /mol_type="other DNA" CDS 1..1563 /codon_start=1 /label=hASCT2-d22 /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSGGVEGMNILGLVVFAIVFG VALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEMEDVGLLFAR LGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTSSSSATLPLM MKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLDFVKIITILV TATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTVLNVEGDALG AGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPAGDATVASEK ESVM" 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 610..615 /label=d22 misc_feature 634..684 /label=TM4c /note="Garaeva et al., 2018" misc_feature 700..816 /label=TM5 /note="Garaeva et al., 2018" misc_feature 829..927 /label=TM6 /note="Garaeva et al., 2018" misc_feature 937..990 /label=HP1a /note="Garaeva et al., 2018" misc_feature 997..1038 /label=HP1b /note="Garaeva et al., 2018" misc_feature 1054..1095 /label=TM7a /note="Garaeva et al., 2018" misc_feature 1102..1149 /label=TM7b /note="Garaeva et al., 2018" misc_feature 1171..1215 /label=HP2a /note="Garaeva et al., 2018" misc_feature 1240..1278 /label=HP2b /note="Garaeva et al., 2018" misc_feature 1303..1422 /label=TM8 /note="Garaeva et al., 2018" CDS 1579..1593 /codon_start=1 /label=GGGGS linker /translation="GGGGS" CDS 1594..2310 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG FVTAAAITHGMDELYK" misc_feature 2328..2914 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 2915..3865 /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 3979..4364 /label=dLTR V_segment complement(4365..7108) /label=pLitmus 28i promoter complement(4420..4438) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(4420..4438) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4459..4475) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4483..4499 /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(4507..4537) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4552..4573 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4634..5222) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" rep_origin 5333..5842 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(5884..6744) /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(6745..6849) /gene="bla" /label=AmpR promoter primer_bind 7070..7086 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7090..7108 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 7090..7108 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" LTR 7109..7500 LTR 7109..7494 /label=dLTR misc_feature 7501..7518 /label=PBS /note="the same as for RCAS" misc_feature 7555..7659 /label=packaging signal /note="according to Banks JD and Linial ML, J. Virol., 2000" gene 7771..8033 /label=p19 /note="p19" gene 8034..8047 /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 tttcgctccg gaggagtcga ggggatgaac atcctgggct tggtagtgtt tgcaattgtc 661 tttggtgtgg cgctgcggaa gctggggcct gaaggggagc tgcttatccg cttcttcaac 721 tccttcaatg aggccaccat ggttctggtc tcctggatca tgtggtacgc ccctgtgggc 781 atcatgttcc tggtggctgg caagatcgtg gagatggagg atgtgggttt actctttgcc 841 cgccttggca agtacattct gtgctgcctg ctgggtcacg ccatccatgg gctcctggta 901 ctgcccctca tctacttcct cttcacccgc aaaaacccct accgcttcct gtggggcatc 961 gtgacgccgc tggccactgc ctttgggacc tcttccagtt ccgccacgct gccgctgatg 1021 atgaagtgcg tggaggagaa taatggcgtg gccaagcaca tcagccgttt catcctgccc 1081 atcggcgcca ccgtcaacat ggacggtgcc gcgctcttcc agtgcgtggc cgcagtgttc 1141 attgcacagc tcagccagca gtccttggac ttcgtaaaga tcatcaccat cctggtcacg 1201 gccacagcgt ccagcgtggg ggcagcgggc atccctgctg gaggtgtcct cactctggcc 1261 atcatcctcg aagcagtcaa cctcccggtc gaccatatct ccttgatcct ggctgtggac 1321 tggctagtcg accggtcctg taccgtcctc aatgtagaag gtgacgctct gggggcagga 1381 ctcctccaaa attacgtgga ccgtacggag tcgagaagca cagagcctga gttgatacaa 1441 gtgaagagtg agctgcccct ggatccgctg ccagtcccca ctgaggaagg aaaccccctc 1501 ctcaaacact atcgggggcc cgcaggggat gccacggtcg cctctgagaa ggaatcagtc 1561 atgactagta ccggtaccgg aggtggtgga tccgtgagca agggcgccga gctgttcacc 1621 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg 1681 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 1741 accggcaagc tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag 1801 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct 1861 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc 1921 gccgaggtga agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat 1981 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat 2041 gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac 2101 aacatcgagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc 2161 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag 2221 gaccccaacg agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc 2281 acccacggca tggatgagct gtacaagtga gcggccgcga ctctagaccc ctctccctcc 2341 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat 2401 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct 2461 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg 2521 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta 2581 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag 2641 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg 2701 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat 2761 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca 2821 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc 2881 ctttgaaaaa cacgatgata atatggccac aaccatggcg gaaggatccg tcgccaggca 2941 gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca 3001 tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc 3061 cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga 3121 ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc 3181 accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg 3241 ccatgatcag ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca 3301 ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga 3361 aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat 3421 gatctatcgc ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga 3481 ggtcgagtca aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc aggcccgtcg 3541 gctctatacc atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt 3601 caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg 3661 cagcgtctcg cccgtccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc 3721 gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc 3781 gtactacgtc gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg ttctggcctg 3841 gcagatcggc gtgatggaag agtgacgcgt atctgagggg actagggtgt gtttaggcga 3901 aaagcggggc ttcggttgta cgcggttagg agtcccctcg aggtatggca gatatgcttt 3961 tgcataggga gggggaaatg tagtcttaat cataggttaa catgtatatt accaaataag 4021 ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg agggagcgac 4081 ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt cctcatcgga 4141 tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg tggcgcatgc 4201 tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg tagttgctaa 4261 caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga tggccggacc 4321 gtcgattccc tgacgactac gagcacctga atgaagcaga aggcatcctg caggaattcc 4381 tcgagacgta cggtgcgcgc gatgcattcg aagatctgcc ctatagtgag tcgtattacg 4441 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4501 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4561 ctaactcaca ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttacgcgccc 5341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 5401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgct 5461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 5521 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat 5581 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 5641 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc 5701 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 5761 acaaaatatt aacgtttaca atttaaatat ttgcttatac aatcttcctg tttttggggc 5821 ttttctgatt atcaaccggg gtaaatcaat ctaaagtata tatgagtaaa cttggtctga 5881 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 5941 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6001 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 6061 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 6121 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 6181 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 6241 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 6301 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 6361 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 6421 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 6481 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt 6541 gctcatcatt ggagaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 6601 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 6661 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 6721 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 6781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 6841 ggttccgcgc acatttcccc gaaaagtgcc acctgacgta gttaacaaaa aaaagcccgc 6901 cgaagcgggc tttattacca agcgaagcgc cattcgccat tcaggctgcg caactgttgg 6961 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct 7021 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 7081 gccagtccgt aatacgactc actataggtg tagtcttaat cataggttaa catgtatatt 7141 accaaataag ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg 7201 agggagcgac ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt 7261 cctcatcgga tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg 7321 tggcgcatgc tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg 7381 tagttgctaa caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga 7441 tggccggacc gtcgattccc tgacgactac gagcacctga atgaagcaga aggcttcatt 7501 tggtgacccc gacgtgatcg ttagggaata gtggtcggcc acaggcggcg tggcgatcct 7561 gtctccatcc gtctcgtcga ccgggaggcg agttcgatga ccctgataga gggggctgcg 7621 gcttagggag gcagaagctg agcaccgtcg gtaggagctc cagggcccgg agcgactgac 7681 ccctgccgag aactcagagg gtcgtcggaa gacggagagt gagcccgacg accaccccag 7741 gcacgtcttt ggtcggcctg cggatcaagc tagcaagccg tcattaaggt gatttcgtcc 7801 gcgtgtaaaa cctattgcgg gaaaatctct ccttctaaga aggaaatagg ggccatgttg 7861 tccctgttac aaaaggaagg gttgctttag tctccctcag atttatattc tccggggtcc 7921 tgggatccca tcactgcggc gctctcccag cgggcaatgg tacttggaaa atcgggagag 7981 ttaaaaacct ggggattggt tttgggggca ttgaaggcgg ctcgagagta acaggactaa 8041 cttgaatcga tgcgctaccg gactcagatc ttgccacc //