LOCUS V043268 8416 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043268 VERSION V043268 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..3270 /label="TACSTD2(NM_002353)" /note="TACSTD2(NM_002353)" /gene="TACSTD2" CDS 3277..3342 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3386..3597 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3610..3878 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3905..4504 /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 4511..5099 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5173..5406 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5478..5599 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5618..5753 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5791..5807) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5815..5831 /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(5839..5869) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5884..5905 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6193..6781) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6952..7812) /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(7813..7917) /gene="bla" /label="AmpR promoter" primer_bind 8391..8407 /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 catggctcgg ggccccggcc tcgcgccgcc accgctgcgg 2341 ctgccgctgc tgctgctggt gctggcggcg gtgaccggcc acacggccgc gcaggacaac 2401 tgcacgtgtc ccaccaacaa gatgaccgtg tgcagccccg acggccccgg cggccgctgc 2461 cagtgccgcg cgctgggctc gggcatggcg gtcgactgct ccacgctgac ctccaagtgt 2521 ctgctgctca aggcgcgcat gagcgccccc aagaacgccc gcacgctggt gcggccgagt 2581 gagcacgcgc tcgtggacaa cgatggcctc tacgaccccg actgcgaccc cgagggccgc 2641 ttcaaggcgc gccagtgcaa ccagacgtcg gtgtgctggt gcgtgaactc ggtgggcgtg 2701 cgccgcacgg acaagggcga cctgagccta cgctgcgatg agctggtgcg cacccaccac 2761 atcctcattg acctgcgcca ccgccccacc gccggcgcct tcaaccactc agacctggac 2821 gccgagctga ggcggctctt ccgcgagcgc tatcggctgc accccaagtt cgtggcggcc 2881 gtgcactacg agcagcccac catccagatc gagctgcggc agaacacgtc tcagaaggcc 2941 gccggtgacg tggatatcgg cgatgccgcc tactacttcg agagggacat caagggcgag 3001 tctctattcc agggccgcgg cggcctggac ttgcgcgtgc gcggagaacc cctgcaggtg 3061 gagcgcacgc tcatctatta cctggacgag attcccccga agttctccat gaagcgcctc 3121 accgccggcc tcatcgccgt catcgtggtg gtcgtggtgg ccctcgtcgc cggcatggcc 3181 gtcctggtga tcaccaaccg gagaaagtcg gggaagtaca agaaggtgga gatcaaggaa 3241 ctgggggagt tgagaaagga accgagcttg ggatccgact acaaagacca tgacggtgat 3301 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3361 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3421 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3481 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3541 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3601 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3661 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3721 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3781 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3841 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3901 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc 3961 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc 4021 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 4081 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 4141 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 4201 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 4261 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 4321 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 4381 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 4441 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 4501 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 4561 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 4621 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 4681 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 4741 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 4801 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 4861 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 4921 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 4981 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5041 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5101 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 5161 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 5221 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 5281 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 5341 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 5401 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 5461 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5521 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5581 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 5641 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 5701 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 5761 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5821 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5881 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5941 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6001 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6061 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6121 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6181 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6241 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6301 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6361 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6421 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6481 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6541 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6601 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 6661 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6721 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6781 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7081 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7921 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 7981 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8041 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8101 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8161 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8221 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 8281 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 8341 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 8401 ggccagtgcc aagctg //