LOCUS V038258 9271 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038258 VERSION V038258 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3924 /label="Src(NM_009271)" /note="Src(NM_009271)" /gene="Src" CDS 3931..3996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4040..4251 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4264..4532 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4565..5359 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5366..5954 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6028..6261 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6333..6454 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6473..6608 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6646..6662) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6670..6686 /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(6694..6724) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6739..6760 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7048..7636) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7807..8667) /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(8668..8772) /gene="bla" /label="AmpR promoter" primer_bind 9246..9262 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgggcagc aacaagagca agcccaagga cgccagccag 2341 cggcgccgca gcctggagcc ctcggaaaac gtgcacgggg cagggggcgc cttcccggcc 2401 tcacagacac cgagcaagcc cgcctccgcc gacggccacc gcgggcccag cgccgccttc 2461 gtgccgcccg cggccgagcc caagctcttc ggaggcttca actcctcgga caccgtcacc 2521 tccccgcaga gggcggggcc tctggcaggt ggggtgacca cctttgtggc cctctatgac 2581 tatgagtcac ggacagagac tgacctgtcc ttcaagaaag gggagcggct gcagattgtc 2641 aataacacga ggaaggtgga tgtcagagag ggagactggt ggctggcaca ctcgctgagc 2701 acgggacaga ccggttacat ccccagcaac tatgtggcgc cctccgactc catccaggct 2761 gaggagtggt actttggcaa gatcactaga cgggaatcag agcggctgct gctcaacgcc 2821 gagaacccga gagggacctt cctcgtgagg gagagtgaga ccacaaaagg tgcctactgc 2881 ctctctgtat ccgacttcga caatgccaag ggcctaaatg tgaaacacta caagatccgc 2941 aagctggaca gcggcggttt ctacatcacc tcccgcaccc agttcaacag cctgcagcag 3001 ctcgtggctt actactccaa acatgctgat ggcctgtgtc accgcctcac taccgtatgt 3061 cccacatcca agcctcagac ccagggattg gccaaggatg cgtgggagat cccccgggag 3121 tccctgcggc tggaggtcaa gctgggccag ggttgcttcg gagaggtgtg gatggggacc 3181 tggaacggca ccacgagggt tgccatcaaa actctgaagc caggcaccat gtccccagag 3241 gccttcctgc aggaggccca agtcatgaag aaactgaggc acgagaaact ggtgcagctg 3301 tatgctgtgg tgtcggaaga acccatttac attgtgacag agtacatgaa caaggggagt 3361 ctgctggact ttctcaaggg ggaaacgggc aaatatttgc ggctacccca gctggtggac 3421 atgtctgctc agatcgcttc aggcatggcc tatgtggagc ggatgaacta tgtgcaccgg 3481 gaccttcgag ccgccaatat cctagtaggg gagaacctgg tgtgcaaagt ggccgacttt 3541 gggttggccc ggctcataga agacaacgaa tacacagccc ggcaaggtgc caaattcccc 3601 atcaagtgga ccgcccctga agctgctctg tacggcaggt tcaccatcaa gtcggatgtg 3661 tggtcctttg ggattctgct gaccgagctc accactaagg gaagagtgcc ctatcctggg 3721 atggtgaacc gtgaggttct ggaccaggtg gagcggggct accggatgcc ttgtcccccc 3781 gagtgccccg agtccctgca tgaccttatg tgccagtgct ggcggaagga gcccgaggag 3841 cggcccacct tcgagtacct gcaggccttc ctggaagact actttacgtc cactgagcca 3901 cagtaccagc ccggggagaa cctaggatcc gactacaaag accatgacgg tgattataaa 3961 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 4021 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 4081 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 4141 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4201 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4261 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4321 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4381 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4441 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4501 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctaggc 4561 caccatggct gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 4621 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 4681 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 4741 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 4801 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 4861 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 4921 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 4981 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 5041 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 5101 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 5161 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 5221 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 5281 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 5341 tcttgacgag ttcttctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 5401 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 5461 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 5521 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 5581 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 5641 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 5701 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 5761 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 5821 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 5881 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 5941 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 6001 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 6061 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6121 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6181 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6241 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6301 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6421 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6481 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6541 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6601 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8881 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8941 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9001 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9061 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9241 acgttgtaaa acgacggcca gtgccaagct g //