LOCUS V036236 9790 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036236 VERSION V036236 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1804..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3816 /label="SLC16A1(NM_003051)" /note="SLC16A1(NM_003051)" /gene="SLC16A1" CDS 3823..3888 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3932..4143 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4156..4424 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4453..4462 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4483..5145 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5215..5268 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5269..5868 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 5869..6457 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6531..6764 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6836..6957 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6997..7132 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7165..7181) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7189..7205 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(7213..7243) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7258..7279 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7567..8155) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8326..9186) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(9187..9291) /gene="bla" /label="AmpR promoter" primer_bind 9765..9781 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgc caccagcagt tggaggtcca 2341 gttggataca cccccccaga tggaggctgg ggctgggcag tggtaattgg agctttcatt 2401 tccatcggct tctcttatgc atttcccaaa tcaattactg tcttcttcaa agagattgaa 2461 ggtatattcc atgccaccac cagcgaagtg tcatggatat cctccataat gttggctgtc 2521 atgtatggtg gaggtcctat cagcagtatc ctggtgaata aatatggaag tcgtatagtc 2581 atgattgttg gtggctgctt gtcaggctgt ggcttgattg cagcttcttt ctgtaacacc 2641 gtacagcaac tatacgtctg tattggagtc attggaggtc ttgggcttgc cttcaacttg 2701 aatccagctc tgaccatgat tggcaagtat ttctacaaga ggcgaccatt ggccaacgga 2761 ctggccatgg caggcagccc tgtgttcctc tgtactctgg cccccctcaa tcaggttttc 2821 ttcggtatct ttggatggag aggaagcttt ctaattcttg ggggcttgct actaaactgc 2881 tgtgttgctg gagccctcat gcgaccaatc gggcccaagc caaccaaggc agggaaagat 2941 aagtctaaag catcccttga gaaagctgga aaatctggtg tgaaaaaaga tctgcatgat 3001 gcaaatacag atcttattgg aagacaccct aaacaagaga aacgatcagt cttccaaaca 3061 attaatcagt tcctggactt aaccctattc acccacagag gctttttgct atacctctct 3121 ggaaatgtga tcatgttttt tggactcttt gcacctttgg tgtttcttag tagttatggg 3181 aagagtcagc attattctag tgagaagtct gccttccttc tttccattct ggcttttgtt 3241 gacatggtag cccgaccatc tatgggactt gtagccaaca caaagccaat aagacctcga 3301 attcagtatt tctttgcggc ttccgttgtt gcaaatggag tgtgtcatat gctagcacct 3361 ttatccacta cctatgttgg attctgtgtc tatgcgggat tctttggatt tgccttcggg 3421 tggctcagct ccgtattgtt tgaaacattg atggaccttg ttggacccca gaggttctcc 3481 agcgctgtgg gattggtgac cattgtggaa tgctgtcctg tcctcctggg gccaccactt 3541 ttaggtcggc tcaatgacat gtatggagac tacaaataca catactgggc atgtggcgtc 3601 gtcctaatta tttcaggtat ctatctcttc attggcatgg gcatcaatta tcgacttttg 3661 gcaaaagaac agaaagcaaa cgagcagaaa aaggaaagta aagaggaaga gaccagtata 3721 gatgttgctg ggaagccaaa tgaagttacc aaagcagcag aatctccgga ccagaaagac 3781 acagatggag ggcccaagga ggaggaaagt ccagtcggat ccgactacaa agaccatgac 3841 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3901 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3961 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 4021 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 4081 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 4141 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4201 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4261 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4321 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4381 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4441 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4501 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4561 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4621 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4681 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4741 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4801 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4861 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4921 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4981 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 5041 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 5101 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 5161 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5221 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5281 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5341 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5401 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5461 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5521 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5581 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5641 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5701 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5761 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5821 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5881 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5941 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 6001 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6061 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6121 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6181 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6241 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6301 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6361 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6421 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6481 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6541 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6781 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6841 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6901 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6961 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 7021 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 7081 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 7141 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7201 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7261 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7321 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7981 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 8041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 9001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9301 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9361 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9421 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9481 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9541 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9601 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9661 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9721 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9781 tgccaagctg //