LOCUS V035368 8504 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035368 VERSION V035368 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..3406 /label="Anxa1(NM_010730)" /note="Anxa1(NM_010730)" /gene="Anxa1" promoter 3474..3685 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3698..3966 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3993..4592 /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 4599..5187 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5261..5494 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5566..5687 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5706..5841 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5879..5895) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5903..5919 /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(5927..5957) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5972..5993 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6281..6869) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7040..7900) /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(7901..8005) /gene="bla" /label="AmpR promoter" primer_bind 8479..8495 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat ggcaatggta tcagaattcc tcaagcaggc 2401 ccgttttctt gaaaatcaag aacaggaata tgttcaagct gtaaaatcat acaaaggtgg 2461 tcctgggtca gcagtgagcc cctacccttc cttcaatgta tcctcggatg ttgctgcctt 2521 gcacaaagct atcatggtta aaggtgtgga tgaagcaacc atcattgaca ttcttaccaa 2581 gaggaccaat gctcagcgcc agcagatcaa ggccgcgtac ttacaggaga atggaaagcc 2641 cttggatgaa gtcttgagaa aagcccttac aggccacctg gaggaggttg ttttggctat 2701 gctaaaaact ccagctcagt ttgatgcaga tgaactccgt ggtgccatga agggacttgg 2761 aacagatgaa gacactctca ttgagatttt gacaacaaga tctaacgaac aaatcagaga 2821 gattaataga gtctacagag aagagctgaa aagagatctg gccaaagaca tcacttcaga 2881 tacatctgga gactttcgga aagccttgct tgctcttgcc aagggtgacc gttgtcagga 2941 cttgagtgtg aatcaagatt tggctgatac agatgccagg gctttgtatg aagctggaga 3001 aaggagaaag gggacagacg tgaacgtctt caccacaatt ctgaccagca ggagctttcc 3061 tcatcttcgc agagtgtttc agaattacgg aaagtacagt caacatgaca tgaacaaagc 3121 tctggatctg gaactgaagg gtgacattga gaagtgcctc acaaccatcg tgaagtgtgc 3181 caccagcact ccagctttct ttgccgagaa gctgtacgaa gccatgaagg gtgccggaac 3241 tcgccataag gcattgatca ggattatggt ctcccgttcg gaaattgaca tgaatgaaat 3301 caaagtattt taccagaaga agtatggaat ctctctttgc caagccatcc tggatgaaac 3361 caaaggagac tatgaaaaaa tcctggtggc tctgtgtggt ggaaactagg gatccctcga 3421 gagatcagtg tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 3481 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 3541 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 3601 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 3661 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 3721 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 3781 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 3841 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 3901 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 3961 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc 4021 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 4081 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 4141 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 4201 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 4261 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 4321 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 4381 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 4441 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 4501 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 4561 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 4621 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 4681 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 4741 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 4801 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 4861 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 4921 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 4981 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 5041 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 5101 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 5161 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5221 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5281 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5341 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5401 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5461 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5521 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 5581 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 5641 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 5701 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 5761 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 5821 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 5881 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5941 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6001 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6721 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8101 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8161 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8221 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8281 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8341 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8401 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8461 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //