LOCUS V032783 8746 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032783 VERSION V032783 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..3600 /label="CCNB1(NM_031966)" /note="CCNB1(NM_031966)" /gene="CCNB1" CDS 3607..3672 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3716..3927 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3940..4208 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4235..4834 /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 4841..5429 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5503..5736 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5808..5929 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5948..6083 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6121..6137) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6145..6161 /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(6169..6199) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6214..6235 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6523..7111) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7282..8142) /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(8143..8247) /gene="bla" /label="AmpR promoter" primer_bind 8721..8737 /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 catggcgctc cgagtcacca ggaactcgaa aattaatgct 2341 gaaaataagg cgaagatcaa catggcaggc gcaaagcgcg ttcctacggc ccctgctgca 2401 acctccaagc ccggactgag gccaagaaca gctcttgggg acattggtaa caaagtcagt 2461 gaacaactgc aggccaaaat gcctatgaag aaggaagcaa aaccttcagc tactggaaaa 2521 gtcattgata aaaaactacc aaaacctctt gaaaaggtac ctatgctggt gccagtgcca 2581 gtgtctgagc cagtgccaga gccagaacct gagccagaac ctgagcctgt taaagaagaa 2641 aaactttcgc ctgagcctat tttggttgat actgcctctc caagcccaat ggaaacatct 2701 ggatgtgccc ctgcagaaga agacctgtgt caggctttct ctgatgtaat tcttgcagta 2761 aatgatgtgg atgcagaaga tggagctgat ccaaaccttt gtagtgaata tgtgaaagat 2821 atttatgctt atctgagaca acttgaggaa gagcaagcag tcagaccaaa atacctactg 2881 ggtcgggaag tcactggaaa catgagagcc atcctaattg actggctagt acaggttcaa 2941 atgaaattca ggttgttgca ggagaccatg tacatgactg tctccattat tgatcggttc 3001 atgcagaata attgtgtgcc caagaagatg ctgcagctgg ttggtgtcac tgccatgttt 3061 attgcaagca aatatgaaga aatgtaccct ccagaaattg gtgactttgc ttttgtgact 3121 gacaacactt atactaagca ccaaatcaga cagatggaaa tgaagattct aagagcttta 3181 aactttggtc tgggtcggcc tctacctttg cacttccttc ggagagcatc taagattgga 3241 gaggttgatg tcgagcaaca tactttggcc aaatacctga tggaactaac tatgttggac 3301 tatgacatgg tgcactttcc tccttctcaa attgcagcag gagctttttg cttagcactg 3361 aaaattctgg ataatggtga atggacacca actctacaac attacctgtc atatactgaa 3421 gaatctcttc ttccagttat gcagcacctg gctaagaatg tagtcatggt aaatcaagga 3481 cttacaaagc acatgactgt caagaacaag tatgccacat cgaagcatgc taagatcagc 3541 actctaccac agctgaattc tgcactagtt caagatttag ccaaggctgt ggcaaaggtg 3601 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat 3661 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca 3721 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt 3781 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 3841 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 3901 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca 3961 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct 4021 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg 4081 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg 4141 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt 4201 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg 4261 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga 4321 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct 4381 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 4441 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 4501 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 4561 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 4621 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 4681 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 4741 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 4801 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa 4861 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 4921 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 4981 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 5041 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 5101 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 5161 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 5221 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 5281 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 5341 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 5401 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 5461 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 5521 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 5581 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 5641 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 5701 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 5761 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 5821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 5941 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 6001 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 6061 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 6121 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6181 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6241 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6961 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8281 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8341 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8401 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8461 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8521 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8581 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 8641 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 8701 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //