LOCUS V018674 9105 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018674 VERSION V018674 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 2827..3831 /label="Larp1b(NM_001014036)" /note="Larp1b(NM_001014036)" /gene="Larp1b" CDS 3838..3861 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3893..4392 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4413..5012 /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 5026..5614 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5821..6454 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6583..6599) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6607..6623 /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(6631..6661) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6676..6697 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6985..7573) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7744..8604) /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(8605..8709) /gene="bla" /label="AmpR promoter" polyA_signal 8757..8891 /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 gccaccatgg caaactggcc aacaccaggt gaattggtta acacggggtc tcagagtgtt 2881 atcagccaag gaaataaaaa gccacaaatt agaaaagaaa aagaagagaa gattgaaaaa 2941 agaagcaaca gtgagagcaa agagaaccgg gaagcaaaac tagatggtcc tattgaaaac 3001 gtcagcgagg atgaagctca gtcaagcagc cagcggaaga gagctaataa gcacaaatgg 3061 gtaccactcc acttagatga tgtgagacca gacagccagg aaagacctgg gtcccggaac 3121 agctctagat gtcaacctga agcaaataaa ccagcgcaca acaatagaaa aagtgataca 3181 cgaagttgga gacgggatag agaaaagaga gatgatcaag atgaagtatc cagtgtgaga 3241 agtgagggtg gtaccatccg aggttccact agaggccggg gaagaggccg gggccgagga 3301 agaggccgag gtcgaggaaa ccctagattg aactttgatt attcatatgg ttatcgagag 3361 ccaggggaaa ggactgagca gccatttcct acagagctta acaccagcat gatgtattac 3421 tatgatgatg gcacaggtgt gcaggtgtat cctgtggaag aaacactgct taaagagtac 3481 attaagcgcc aaattgagta ttacttcagc atagagaact tggaacggga cttctttctt 3541 cgaagaaaga tggatgagca gggatttctg cccatttccc tgattgctgg ttttcaccgt 3601 gtgcaggctc tcaccacaaa ccttaatctc atcttggagg cacttaagga tagcacagaa 3661 gtagaaattg tggatgagaa aatgaggaaa aagatagaac cagaaaaatg gccaattcca 3721 ggccctcctc cacgaaatgt gccacagaca gacttctctc agttgattga ttgtccggag 3781 ttcatacctg gccaggcctt tggctcccat acagtcaggg tgatgatcta cgaattcgat 3841 tacaaggatg acgacgataa gtgaggatcc cgcgactcta gataattcta ccgggtaggg 3901 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 3961 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 4021 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 4081 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 4141 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 4201 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 4261 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 4321 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 4381 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 4441 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 4501 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 4561 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 4621 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 4681 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 4741 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 4801 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 4861 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 4921 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 4981 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat 5041 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5101 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5161 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5221 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5281 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5341 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5401 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 5461 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5521 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5581 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 5641 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 5701 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 5761 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 5821 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 5881 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 5941 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 6001 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 6061 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 6121 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 6181 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 6241 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 6301 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6361 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6421 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6481 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 6541 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 6601 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 6661 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 6721 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 6781 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6841 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6901 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6961 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 7021 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 7081 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 7141 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 7201 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 7261 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 7321 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 7381 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 7441 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 7501 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 7561 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 7621 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 7681 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 7741 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 7801 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 7861 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 7921 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 7981 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 8041 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 8101 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 8161 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 8221 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 8281 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 8341 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 8401 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 8461 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 8521 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 8581 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 8641 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 8701 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact 8761 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 8821 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 8881 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat 8941 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg 9001 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt 9061 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //