LOCUS V045564 9431 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045564 VERSION V045564 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..3525 /label="Krt19(NM_008471)" /note="Krt19(NM_008471)" /gene="Krt19" promoter 3590..3801 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3814..4082 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4111..4120 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4141..4803 /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 4873..4926 /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 4927..5526 /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 5527..6114 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6188..6421 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6493..6614 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6633..6768 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6806..6822) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6830..6846 /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(6854..6884) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6899..6920 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7208..7796) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7967..8827) /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(8828..8932) /gene="bla" /label="AmpR promoter" primer_bind 9406..9422 /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 gccaccatga cttcctatag ctatcgccag 2341 acctcagcta tgtcttcctt tgggggtacg ggcgggggtt cagtacgcat tgggtcaggg 2401 ggtgttttcc gcgcacccag catccacggg ggctcaggtg gccgcggcgt gtccgtgtcc 2461 tccacccgct tcgtgacctc gtcctccggg agctatggcg gagtccgcgg tggaagtttt 2521 agtgggaccc tggctgtgtc tgatgggctg ctgtctggca atgagaagat caccatgcaa 2581 aacctcaatg atcgtctcgc ctcctacttg gacaaggtgc gcgccctaga gcaggccaat 2641 ggcgagctgg aggtgaagat ccgcgactgg taccagaagc agggacccgg accctcccga 2701 gattacaacc actactttaa gaccatcgag gacttgcgcg acaagattct tggtgccacc 2761 attgacaact ccaagattgt cctacagatt gacaatgctc gcctggctgc agatgacttc 2821 agaaccaagt ttgagacaga acacgccttg cgtctgagcg tggaggccga catcaacggc 2881 ctgcgccggg tgctggatga gctgactctg gccaggactg acctggagat gcagattgag 2941 agcctgaagg aggagctggc ctacctgaag aagaaccatg aggaggaaat tactgccctg 3001 aggagccagg tgggtggcca ggtcagtgtg gaggtggatt ccactcccgg tgtcgaccta 3061 gccaagatcc tgagtgagat gagaagtcag tatgagatca tggccgagaa gaaccggaag 3121 gatgctgaag ccacctacct tgctcggatt gaggagctga acacccaggt cgccgtccac 3181 tctgagcaga tccagataag caagaccgaa gtcacggacc ttcgacggac cctccagggc 3241 cttgagattg agctgcagtc ccagctcagc atgaaagctg ccctggaagg cacgctggca 3301 gagacggagg cccgttatgg agtccagctg tcacagatcc agagcgtgat cagcggtttt 3361 gaagcccagc tgagcgacgt gcgtgccgac atagagcgcc agaaccagga gtataagcag 3421 ctcatggaca tcaagtccag gctggagcag gagatcgcca cctaccgcag cctgctggag 3481 ggccaggaag cccactacaa caatctgccc acccccaagg ccatctgagg atccctgcga 3541 tatccagcgc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc 3601 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag 3661 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc 3721 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac 3781 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc 3841 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc 3901 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg 3961 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc 4021 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga 4081 tccaagctgt gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg 4141 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg 4201 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc 4261 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac 4321 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc 4381 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc 4441 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc 4501 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac 4561 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc 4621 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac 4681 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac 4741 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc 4801 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc 4861 accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc 4921 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 4981 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 5041 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 5101 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 5161 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 5221 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 5281 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 5341 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 5401 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 5461 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 5521 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 5581 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 5641 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 5701 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 5761 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 5821 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 5881 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 5941 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 6001 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 6061 cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta 6121 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 6181 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact 6241 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 6301 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 6361 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 6421 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 6481 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 6601 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg 6661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 6721 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca 6781 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 6841 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 6901 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 6961 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 7021 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 7081 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 7141 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 7201 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 7261 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7321 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7381 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7441 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7501 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7561 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7621 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 7681 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7741 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7801 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7861 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 7921 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 7981 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 8041 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 8101 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 8161 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 8221 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 8281 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8341 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 8401 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8461 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8521 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8581 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8641 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8701 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8761 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 8821 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 8881 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 8941 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 9001 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 9061 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 9121 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 9181 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 9241 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 9301 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 9361 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 9421 gtgccaagct g //