LOCUS V047224 8131 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047224 VERSION V047224 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 2323..2985 /label="RAB27A(NM_004580)" /note="RAB27A(NM_004580)" /gene="RAB27A" CDS 2992..3057 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3101..3312 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3325..3593 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3620..4219 /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 4226..4814 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4888..5121 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5193..5314 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5333..5468 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5506..5522) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5530..5546 /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(5554..5584) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5599..5620 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5908..6496) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6667..7527) /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(7528..7632) /gene="bla" /label="AmpR promoter" primer_bind 8106..8122 /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 catgccgaag aagaagcgcg tgatgtctga tggagattat 2341 gattacctca tcaagttttt agctttggga gactctggtg tagggaagac cagtgtactt 2401 taccaatata cagatggtaa atttaactcc aaatttatca caacagtggg cattgatttc 2461 agggaaaaaa gagtggtgta cagagccagt gggccggatg gagccactgg cagaggccag 2521 agaatccacc tgcagttatg ggacacagca gggcaggaga ggtttcgtag cttaacgaca 2581 gcgttcttca gagatgctat gggttttctt ctactttttg atctgacaaa tgagcaaagt 2641 ttcctcaatg tcagaaactg gataagccag ctacagatgc atgcatattg tgaaaaccca 2701 gatatagtgc tgtgtggaaa caagagtgat ctggaggacc agagagtagt gaaagaggag 2761 gaagccatag cactcgcaga gaaatatgga atcccctact ttgaaactag tgctgccaat 2821 gggacaaaca taagccaagc aattgagatg cttctggacc tgataatgaa gcgaatggaa 2881 cggtgtgtgg acaagtcctg gattcctgaa ggagtggtgc gatcaaatgg tcatgcctct 2941 acggatcagt taagtgaaga aaaggagaaa ggggcatgtg gctgtggatc cgactacaaa 3001 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3061 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3121 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3181 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3241 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3301 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3361 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3421 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 3481 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 3541 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 3601 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 3661 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 3721 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 3781 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 3841 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 3901 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 3961 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4021 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4081 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4141 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 4201 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 4261 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 4321 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 4381 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 4441 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 4501 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 4561 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 4621 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 4681 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 4741 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 4801 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 4861 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 4921 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 4981 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5041 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5101 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 5161 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 5341 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 5401 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 5461 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 5521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 5581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 5641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 5701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 5761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 5821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 5881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 6661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 7561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 7621 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 7681 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 7741 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 7801 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 7861 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 7921 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 7981 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8041 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8101 acgttgtaaa acgacggcca gtgccaagct g //