LOCUS V030698 9636 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030698 VERSION V030698 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 2301..4490 /label="DTL(NM_016448)" /note="DTL(NM_016448)" /gene="DTL" CDS 4497..4562 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4606..4817 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4830..5098 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5125..5724 /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 5731..6319 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6393..6626 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6698..6819 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6838..6973 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7011..7027) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7035..7051 /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(7059..7089) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7104..7125 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7413..8001) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8172..9032) /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(9033..9137) /gene="bla" /label="AmpR promoter" primer_bind 9611..9627 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atgctcttca attcggtgct ccgccagccc cagcttggcg 2341 tcctgagaaa tggatggtct tcacaatacc ctcttcaatc ccttctgact ggttatcagt 2401 gcagtggtaa tgatgaacac acttcttatg gagaaacagg agtcccagtt cctccttttg 2461 gatgtacctt ctcttctgct cccaatatgg aacatgtact agcagttgcc aatgaagaag 2521 gctttgttcg attgtataac acagaatcac aaagtttcag aaagaagtgc ttcaaagaat 2581 ggatggctca ctggaatgcc gtctttgacc tggcctgggt tcctggtgaa cttaaacttg 2641 ttacagcagc aggtgatcaa acagccaaat tttgggacgt aaaagctggt gagctgattg 2701 gaacatgcaa aggtcatcaa tgcagcctca agtcagttgc cttttctaag tttgagaaag 2761 ctgtattctg tacgggtgga agagatggca acattatggt ctgggatacc aggtgcaaca 2821 aaaaagatgg gttttatagg caagtgaatc aaatcagtgg agctcacaat acctcagaca 2881 agcaaacccc ttcaaaaccc aagaagaaac agaattcaaa aggacttgct ccttctgtgg 2941 atttccagca aagtgttact gtggtcctct ttcaagacga gaatacctta gtctcagcag 3001 gagctgtgga tgggataatc aaagtatggg atttacgtaa gaattatact gcttatcgac 3061 aagaacccat agcatccaag tctttcctgt acccaggtag cagcactcga aaacttggat 3121 attcaagtct gattttggat tccactggct ctactttatt tgctaattgc acagacgata 3181 acatctacat gtttaatatg actgggttga agacttctcc agtggctatt ttcaatggac 3241 accagaactc taccttttat gtaaaatcca gccttagtcc agatgaccag tttttagtca 3301 gtggctcaag tgatgaagct gcctacatat ggaaggtctc cacaccctgg caacctccta 3361 ctgtgctcct gggtcattct caagaggtca cgtctgtgtg ctggtgtcca tctgacttca 3421 caaagattgc tacctgttct gatgacaata cactaaaaat ctggcgcttg aatagaggct 3481 tagaggagaa accaggaggt gataaacttt ccacggtggg ttgggcctct cagaagaaaa 3541 aagagtcaag acctggccta gtaacagtaa cgagtagcca gagtactcct gccaaagccc 3601 ccagggcaaa gtgcaatcca tccaattctt ccccgtcatc cgcagcttgt gccccaagct 3661 gtgctggaga cctccctctt ccttcaaata ctcctacgtt ctctattaaa acctctcctg 3721 ccaaggcccg gtctcccatc aacagaagag gctctgtctc ctccgtctct cccaagccac 3781 cttcatcttt caagatgtcg attagaaact gggtgacccg aacaccttcc tcatcaccac 3841 ccatcactcc acctgcttcg gagaccaaga tcatgtctcc gagaaaagcc cttattcctg 3901 tgagccagaa gtcatcccaa gcagaggctt gctctgagtc tagaaataga gtaaagagga 3961 ggctagactc aagctgtctg gagagtgtga aacaaaagtg tgtgaagagt tgtaactgtg 4021 tgactgagct tgatggccaa gttgaaaatc ttcatttgga tctgtgctgc cttgctggta 4081 accaggaaga ccttagtaag gactctctag gtcctaccaa atcaagcaaa attgaaggag 4141 ctggtaccag tatctcagag cctccgtctc ctatcagtcc gtatgcttca gaaagctgtg 4201 gaacgctacc tcttcctttg agaccttgtg gagaagggtc tgaaatggta ggcaaagaga 4261 atagttcccc agagaataaa aactggttgt tggccatggc agccaaacgg aaggctgaga 4321 atccatctcc acgaagtccg tcatcccaga cacccaattc caggagacag agcggaaaga 4381 aattgccaag cccggtcacc atcacgccca gctccatgag gaaaatctgc acatacttcc 4441 atagaaagtc ccaggaggac ttctgtggtc ctgaacactc aacagaatta ggatccgact 4501 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca 4561 agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat 4621 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa 4681 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 4741 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 4801 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc 4861 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg 4921 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc 4981 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga 5041 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca 5101 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc 5161 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5221 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5281 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5341 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5401 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5461 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5521 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5581 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5641 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5701 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa 5761 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 5821 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 5881 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 5941 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 6001 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 6061 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 6121 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 6181 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 6241 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 6301 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga 6361 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa 6421 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 6481 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 6541 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 6601 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 6661 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa 6721 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 6781 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc 6841 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6901 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6961 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg 7021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 7081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 7141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 7321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 7381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 7441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 7501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 7561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 7621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 7681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 7741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 7801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 7861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 7921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 7981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 8041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 8101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 8161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 8281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 8341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 8401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 8461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 8521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 8581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 8641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 8701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 8761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 8821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 8881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 8941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 9121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt 9181 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt 9241 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 9301 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 9361 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 9421 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca 9481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 9541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 9601 tcacgacgtt gtaaaacgac ggccagtgcc aagctg //