LOCUS V046352 8971 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046352 VERSION V046352 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..3825 /label="P4HB(NM_000918)" /note="P4HB(NM_000918)" /gene="P4HB" CDS 3832..3897 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3941..4152 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4165..4433 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4460..5059 /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 5066..5654 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5728..5961 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6033..6154 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6173..6308 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6346..6362) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6370..6386 /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(6394..6424) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6439..6460 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6748..7336) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7507..8367) /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(8368..8472) /gene="bla" /label="AmpR promoter" primer_bind 8946..8962 /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 catgctgcgc cgcgctctgc tgtgcctggc cgtggccgcc 2341 ctggtgcgcg ccgacgcccc cgaggaggag gaccacgtcc tggtgctgcg gaaaagcaac 2401 ttcgcggagg cgctggcggc ccacaagtac ctgctggtgg agttctatgc cccttggtgt 2461 ggccactgca aggctctggc ccctgagtat gccaaagccg ctgggaagct gaaggcagaa 2521 ggttccgaga tcaggttggc caaggtggac gccacggagg agtctgacct ggcccagcag 2581 tacggcgtgc gcggctatcc caccatcaag ttcttcagga atggagacac ggcttccccc 2641 aaggaatata cagctggcag agaggctgat gacatcgtga actggctgaa gaagcgcacg 2701 ggcccggctg ccaccaccct gcctgacggc gcagctgcag agtccttggt ggagtccagc 2761 gaggtggctg tcatcggctt cttcaaggac gtggagtcgg actctgccaa gcagtttttg 2821 caggcagcag aggccatcga tgacatacca tttgggatca cttccaacag tgacgtgttc 2881 tccaaatacc agctcgacaa agatggggtt gtcctcttta agaagtttga tgaaggccgg 2941 aacaactttg aaggggaggt caccaaggag aacctgctgg actttatcaa acacaaccag 3001 ctgccccttg tcatcgagtt caccgagcag acagccccga agatttttgg aggtgaaatc 3061 aagactcaca tcctgctgtt cttgcccaag agtgtgtctg actatgacgg caaactgagc 3121 aacttcaaaa cagcagccga gagcttcaag ggcaagatcc tgttcatctt catcgacagc 3181 gaccacaccg acaaccagcg catcctcgag ttctttggcc tgaagaagga agagtgcccg 3241 gccgtgcgcc tcatcaccct ggaggaggag atgaccaagt acaagcccga atcggaggag 3301 ctgacggcag agaggatcac agagttctgc caccgcttcc tggagggcaa aatcaagccc 3361 cacctgatga gccaggagct gccggaggac tgggacaagc agcctgtcaa ggtgcttgtt 3421 gggaagaact ttgaagacgt ggcttttgat gagaaaaaaa acgtctttgt ggagttctat 3481 gccccatggt gtggtcactg caaacagttg gctcccattt gggataaact gggagagacg 3541 tacaaggacc atgagaacat cgtcatcgcc aagatggact cgactgccaa cgaggtggag 3601 gccgtcaaag tgcacagctt ccccacactc aagttctttc ctgccagtgc cgacaggacg 3661 gtcattgatt acaacgggga acgcacgctg gatggtttta agaaattcct ggagagcggt 3721 ggccaggatg gggcagggga tgatgacgat ctcgaggacc tggaagaagc agaggagcca 3781 gacatggagg aagacgatga tcagaaagct gtgaaagatg aactgggatc cgactacaaa 3841 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3901 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3961 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 4021 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 4081 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 4141 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 4201 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 4261 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4321 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4381 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4441 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 4501 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 4561 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 4621 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 4681 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 4741 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 4801 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4861 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4921 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4981 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 5041 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 5101 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 5161 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 5221 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 5281 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 5341 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 5401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 5461 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 5521 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 5581 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 5641 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 5701 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 5761 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5821 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5881 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5941 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6001 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6241 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6301 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6361 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6421 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8461 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8521 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8581 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8641 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8701 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8761 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8821 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8881 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8941 acgttgtaaa acgacggcca gtgccaagct g //