LOCUS V030268 8596 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030268 VERSION V030268 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..3249 /label="BRCC3(NM_024332)" /note="BRCC3(NM_024332)" /gene="BRCC3" CDS 3256..3321 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3365..3576 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3589..3857 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3890..4684 /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 4691..5279 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5353..5586 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5658..5779 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5798..5933 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5971..5987) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5995..6011 /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(6019..6049) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6064..6085 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6373..6961) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7132..7992) /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(7993..8097) /gene="bla" /label="AmpR promoter" primer_bind 8571..8587 /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 catggcggtg caggtggtgc aggcggtgca ggcggttcat 2341 ctcgagtctg acgctttcct cgtttgtctc aaccacgctc tgagcacaga gaaggaggaa 2401 gtaatggggc tgtgcatagg ggagttgaac gatgatacaa ggagtgactc caaatttgca 2461 tatactggaa ctgaaatgcg cacagttgct gaaaaggttg atgccgtcag aattgttcac 2521 attcattctg tcatcatctt acgacgttct gataagagga aggaccgagt agaaatttct 2581 ccagagcagc tgtctgcagc ttcaacagag gcagagaggt tggctgaact gacaggccgc 2641 cccatgagag ttgtgggctg gtatcattcc catcctcata taactgtttg gccttcacat 2701 gttgatgttc gcacacaagc catgtaccag atgatggatc aaggctttgt aggacttatt 2761 ttttcctgtt tcatagaaga taagaacaca aagactggcc gggtactcta cacttgcttc 2821 caatccatac aggcccaaaa gagttcagag tcccttcatg gtccacgaga cttctggagc 2881 tccagccagc acatctccat tgagggccag aaggaagagg aaaggtatga gagaatcgaa 2941 atcccaatcc atattgtacc tcatgtcact atcgggaaag tgtgccttga atcagcagta 3001 gagctgccca agatcctgtg ccaggaggag caggatgcgt ataggaggat ccacagcctt 3061 acacatctgg actcagtaac caagatccat aatggctcag tgtttaccaa gaatctgtgc 3121 agtcagatgt cggcagtcag cgggcctctc ctacagtggt tggaggacag actggagcaa 3181 aaccaacagc atttgcagga attacaacaa gaaaaggaag agcttatgca agaactttct 3241 tctctagaac tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac 3301 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt 3361 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat 3421 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg 3481 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac 3541 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc 3601 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt 3661 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag 3721 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg 3781 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt 3841 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc taggccacca tggctgaaca 3901 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3961 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 4021 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 4081 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4141 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4201 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4261 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4321 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4381 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4441 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4501 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4561 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4621 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4681 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 4741 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 4801 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 4861 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 4921 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 4981 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5041 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 5101 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5161 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5221 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5281 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 5341 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 5401 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 5461 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 5521 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 5581 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 5641 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5701 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5761 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 5821 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 5881 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 5941 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 6001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6781 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8101 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8161 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8221 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8281 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8341 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8401 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 8461 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 8521 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 8581 ggccagtgcc aagctg //