LOCUS V028049 7771 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028049 VERSION V028049 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..2604 /label="SUMO1(NM_003352)" /note="SUMO1(NM_003352)" /gene="SUMO1" CDS 2611..2637 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2641..2667 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2671..2697 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 2741..2952 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2965..3233 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3260..3859 /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 3866..4454 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4528..4761 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4833..4954 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 4973..5108 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5146..5162) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5170..5186 /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(5194..5224) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5239..5260 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5548..6136) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6307..7167) /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(7168..7272) /gene="bla" /label="AmpR promoter" primer_bind 7746..7762 /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 catgtctgac caggaggcaa aaccttcaac tgaggacttg 2341 ggggataaga aggaaggtga atatattaaa ctcaaagtca ttggacagga tagcagtgag 2401 attcacttca aagtgaaaat gacaacacat ctcaagaaac tcaaagaatc atactgtcaa 2461 agacagggtg ttccaatgaa ttcactcagg tttctctttg agggtcagag aattgctgat 2521 aatcatactc caaaagaact gggaatggag gaagaagatg tgattgaagt ttatcaggaa 2581 caaacggggg gtcattcaac agttggatcc tacccatacg acgtacctga ttatgcaact 2641 tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga ctacgcatag 2701 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 2761 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 2821 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 2881 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 2941 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3001 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3061 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 3121 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 3181 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 3241 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 3301 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 3361 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 3421 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 3481 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 3541 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 3601 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 3661 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 3721 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 3781 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 3841 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt 3901 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 3961 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 4021 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 4081 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 4141 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 4201 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 4261 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 4321 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 4381 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 4441 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 4501 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 4561 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 4621 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 4681 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 4741 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 4801 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 4861 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4921 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 4981 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 5041 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 5101 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 5161 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 5221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 5281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 7261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 7321 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 7381 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 7441 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 7501 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 7561 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 7621 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 7681 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 7741 acgttgtaaa acgacggcca gtgccaagct g //