LOCUS V034894 8731 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034894 VERSION V034894 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..3585 /label="RNF128(NM_194463)" /note="RNF128(NM_194463)" /gene="RNF128" CDS 3592..3657 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3701..3912 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3925..4193 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4220..4819 /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 4826..5414 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5488..5721 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5793..5914 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5933..6068 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6106..6122) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6130..6146 /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(6154..6184) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6199..6220 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6508..7096) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7267..8127) /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(8128..8232) /gene="bla" /label="AmpR promoter" primer_bind 8706..8722 /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 catggggccg ccgcctgggg ccggggtctc ctgccgcggt 2341 ggctgcggct tttccagatt gctggcatgg tgcttcctgc tggccctgag tccgcaggca 2401 cccggttccc ggggggctga agcagtgtgg accgcgtacc tcaacgtgtc ctggcgggtt 2461 ccgcacacgg gagtgaaccg tacggtgtgg gagctgagcg aggagggcgt gtacggccag 2521 gactcgccgc tggagcctgt ggctggggtc ctggtaccgc ccgacgggcc cggggcgctt 2581 aacgcctgta acccgcacac gaatttcacg gtgcccacgg tttggggaag caccgtgcaa 2641 gtctcttggt tggccctcat ccaacgcggc gggggctgca ccttcgcaga caagatccat 2701 ctggcttatg agagaggggc gtctggagcc gtcatcttta acttccccgg gacccgcaat 2761 gaggtcatcc ccatgtctca cccgggtgca gtagacattg ttgcaatcat gatcggcaat 2821 ctgaaaggca caaaaattct gcaatctatt caaagaggca tacaagtgac aatggtcata 2881 gaagtaggga aaaaacatgg cccttgggtg aatcactatt caattttttt cgtttctgtg 2941 tcctttttta ttattacggc ggcaactgtg ggctatttta tcttttattc tgctcgaagg 3001 ctacggaatg caagagctca aagcaggaag cagaggcaat taaaggcaga tgctaaaaaa 3061 gctattggaa ggcttcaact acgcacactg aaacaaggag acaaggaaat tggccctgat 3121 ggagatagtt gtgctgtgtg cattgaattg tataaaccaa atgatttggt acgcatctta 3181 acgtgcaacc atattttcca taagacatgt gttgacccat ggctgttaga acacaggact 3241 tgccccatgt gcaaatgtga catactcaaa gctttgggaa ttgaggtgga tgttgaagat 3301 ggatcagtgt ctttacaagt ccctgtatcc aatgaaatat ctaatagtgc ctcctcccat 3361 gaagaggata atcgcagcga gaccgcatca tctggatatg cttcagtaca gggaacagat 3421 gaaccgcctc tggaggaaca cgtgcagtca acaaatgaaa gtctacagct ggtaaaccat 3481 gaagcaaatt ctgtggcagt ggatgttatt cctcatgttg acaacccaac ctttgaagaa 3541 gacgaaactc ctaatcaaga gactgctgtt cgagaaatta aatctggatc cgactacaaa 3601 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3661 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3721 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3781 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3841 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3901 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3961 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 4021 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4081 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4141 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4201 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 4261 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 4321 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 4381 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 4441 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 4501 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 4561 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4621 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4681 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4741 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 4801 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 4861 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 4921 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 4981 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 5041 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 5101 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 5161 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 5221 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 5281 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 5341 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 5401 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 5461 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 5521 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5581 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5641 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5701 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 5761 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 5821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 5941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6001 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6061 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8581 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8641 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8701 acgttgtaaa acgacggcca gtgccaagct g //