LOCUS V031586 10037 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031586 VERSION V031586 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 2303..3850 /label="TRIM6(NM_001003818)" /note="TRIM6(NM_001003818)" /gene="TRIM6" CDS 3857..3946 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 3990..4201 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4214..4482 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4511..4520 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4541..5203 /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 5273..5326 /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 5333..6127 /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 6134..6722 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6794..7027 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7099..7220 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7239..7374 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7412..7428) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7436..7452 /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(7460..7490) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7505..7526 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7814..8402) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8573..9433) /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(9434..9538) /gene="bla" /label="AmpR promoter" primer_bind 10012..10028 /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 gaattcgcca ccatgtgcgg gtcagagagg attctacagg caggaaacat 2341 cttagaaatc agggttgggc aggcaggagc caggagagta gctacaatga cttcaccagt 2401 actggtggac atacgagaag aggtgacctg ccctatctgc ctggagctcc taacagaacc 2461 cctgagcata gactgtggcc acagcttctg ccaagcctgc atcacaccaa atggcaggga 2521 atcagtgatt ggtcaagaag gggaaagaag ctgccctgtg tgccagacca gctaccagcc 2581 agggaacctg cggcctaatc ggcatctggc caacatagtg aggcggctca gagaggtagt 2641 gttgggccct gggaagcagc tgaaagcagt tctttgtgca gaccatggag aaaaactgca 2701 gctcttctgt caggaggatg ggaaggtcat ttgctggctt tgtgagcggt ctcaggagca 2761 ccgtggtcac cacacgttcc tcgtggagga ggttgcccag gagtaccagg agaagtttca 2821 ggagtctcta aagaagctga agaacgagga gcaggaagct gagaagctaa cagcttttat 2881 cagagagaag aaaacatcct ggaagaatca gatggagcct gagagatgca ggatccagac 2941 agagtttaat cagctgcgaa atatcctaga cagagtggag caacgggagc tgaaaaagct 3001 ggaacaggaa gagaagaagg ggctacgaat tatagaagag gctgagaatg atctggtcca 3061 ccagacccag tcgctgcgag agctcatctc ggatctggag cgtcgatgtc aggggtcaac 3121 aatggagctg ctgcaggatg tgagtgatgt cacagaaagg agtgagttct ggaccctgag 3181 gaagccagaa gctctcccta caaagctgag aagtatgttc cgagccccag atctgaaaag 3241 gatgctgcga gtgtgtagag agctgacaga tgtccaaagc tactgggttg acgtgaccct 3301 gaatccacac acagctaatt taaatcttgt cctggctaaa aaccggagac aagtgaggtt 3361 tgtgggagct aaagtatctg gaccttcctg tctggaaaag cattatgact gtagtgtcct 3421 gggctcccag cacttctcct ctggtaagca ttactgggag gtagatgtgg ccaagaagac 3481 tgcctggatc ctgggggtat gcagcaattc actgggacct acattctctt tcaaccattt 3541 tgctcaaaat cacagtgctt actccaggta tcagcctcag agtggatact gggtgattgg 3601 gttacagcat aaccatgaat atagggccta tgaggattct tccccttccc tgcttctctc 3661 catgacagtg ccccctcgcc gtgttggggt tttcttagat tatgaggctg gtactgtctc 3721 cttttataat gtcacaaacc atggcttccc catctacact ttctctaaat attactttcc 3781 cactactctt tgtccatatt ttaatccttg caactgtgta attcctatga ccctgcgtcg 3841 tccaagctct ggatccgaac aaaaactaat atcagaggaa gatttggaac agaaactaat 3901 cagtgaagaa gatttagagc aaaagttaat ttccgaggag gacttatagc ggccgcgaag 3961 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga 4021 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa 4081 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta 4141 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca 4201 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg 4261 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc 4321 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc 4381 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact 4441 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc 4501 tacgctagac gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg 4561 ccgcatcacc ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac 4621 ccccaagcag ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag 4681 cccctacctg ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag 4741 cggctacgag aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat 4801 cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg 4861 ccgcgtgatc ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt 4921 caccgacaag atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa 4981 cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt 5041 cgtggtggac agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg 5101 gggccccatg ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat 5161 cgtggagtac cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc 5221 gtccaattct gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag 5281 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctgcca ccatggctga 5341 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 5401 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 5461 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 5521 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 5581 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 5641 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 5701 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 5761 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 5821 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 5881 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5941 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 6001 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 6061 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 6121 cttctgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 6181 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 6241 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 6301 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 6361 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 6421 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 6481 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 6541 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 6601 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 6661 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 6721 gcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 6781 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct gctttttgct 6841 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6901 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6961 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 7021 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 7081 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 7141 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 7201 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 7261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 7321 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact 7381 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 8041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8221 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 8281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 8341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 8401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 9001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 9301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 9361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9541 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 9601 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 9661 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 9721 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 9781 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 9841 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt 9901 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 9961 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 10021 cggccagtgc caagctg //