LOCUS V040295 8944 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040295 VERSION V040295 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..3700 /label="RNASEH1(NM_002936)" /note="RNASEH1(NM_002936)" /gene="RNASEH1" promoter 3732..4231 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4252..4851 /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 4865..5453 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5660..6293 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6422..6438) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6446..6462 /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(6470..6500) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6515..6536 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6824..7412) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7583..8443) /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(8444..8548) /gene="bla" /label="AmpR promoter" polyA_signal 8596..8730 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatgagctg gcttctgttc ctggcccaca gagtcgcctt 2881 ggccgccttg ccctgccgcc gcggctctcg cgggttcggg atgttctatg ccgtgaggag 2941 gggccgcaag accggggtct ttctgacctg gaatgagtgc agagcacagg tggaccggtt 3001 tcctgctgcc agatttaaga agtttgccac agaggatgag gcctgggcct ttgtcaggaa 3061 atctgcaagc ccggaagttt cagaagggca tgaaaatcaa catggacaag aatcggaggc 3121 gaaagccagc aagcgactcc gtgagccact ggatggagat ggacatgaaa gcgcagagcc 3181 gtatgcaaag cacatgaagc cgagcgtgga gccggcgcct ccagttagca gagacacgtt 3241 ttcctacatg ggagacttcg tcgtcgtcta cactgatggc tgctgctcca gtaatgggcg 3301 tagaaggccg cgagcaggaa tcggcgttta ctgggggcca ggccatcctt taaatgtagg 3361 cattagactt cctgggcggc agacaaacca aagagcggaa attcatgcag cctgcaaagc 3421 cattgaacaa gcaaagactc aaaacatcaa taaactggtt ctgtatacag acagtatgtt 3481 tacgataaat ggtataacta actgggttca aggttggaag aaaaatgggt ggaagacaag 3541 tgcagggaaa gaggtgatca acaaagagga ctttgtggca ctggagaggc ttacccaggg 3601 gatggacatt cagtggatgc atgttcctgg tcattcggga tttataggca atgaagaagc 3661 tgacagatta gccagagaag gagctaaaca atcggaagac tgaggatccc gcgactctag 3721 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 3781 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 3841 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 3901 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 3961 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 4021 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 4081 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 4141 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 4201 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4621 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 4861 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5281 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5461 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5521 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5581 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5641 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5701 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5761 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 5821 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 5881 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 5941 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 6001 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6061 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6121 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6181 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6241 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6301 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6361 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6421 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6481 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6541 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6601 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6661 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6721 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7261 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7381 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7441 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7501 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7561 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7621 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7681 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7741 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7801 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7861 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7921 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 7981 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 8041 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8101 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8161 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8221 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8281 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8341 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8401 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8461 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8521 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 8581 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8641 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8701 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 8761 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 8821 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 8881 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 8941 tagt //