LOCUS V021756 9124 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021756 VERSION V021756 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..3559 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced blue variant of GFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EBFP" CDS 3593..3883 /codon_start=1 /gene="SEC61B" /product="human Sec61β subunit of the translocon in the endoplasmic reticulum" /protein_id="" /transl_table=1 /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS*" /label="Sec61β" promoter 3912..4411 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4432..5031 /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 5045..5633 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5840..6473 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6602..6618) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6626..6642 /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(6650..6680) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6695..6716 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7004..7592) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7763..8623) /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(8624..8728) /gene="bla" /label="AmpR promoter" polyA_signal 8776..8910 /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 ccatggtgag caagggcgag gagctgttca ccggggtggt 2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc cacggcgtgc agtgcttcag 3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 3241 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat 3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3541 catggacgag ctgtacaagt ccggactcag atctcgagga ggagggggaa gcatgcctgg 3601 tccgaccccc agtggcacta acgtgggatc ctcagggcgc tctcccagca aagcagtggc 3661 cgcccgggcg gcgggatcca ctgtccggca gaggaaaaat gccagctgtg ggacaaggag 3721 tgcaggccgc acaacctcgg caggcaccgg ggggatgtgg cgattctaca cagaagattc 3781 acctgggctc aaagttggcc ctgttccagt attggttatg agtcttctgt tcatcgcttc 3841 tgtatttatg ttgcacattt ggggcaagta cactcgttcg tagggatccc gcgactctag 3901 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 3961 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 4021 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 4081 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 4141 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 4201 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 4261 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 4321 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 4381 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4441 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4501 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4561 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4621 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4681 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4741 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4801 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4861 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4921 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4981 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 5041 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5101 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5161 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5221 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5281 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5341 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5401 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5461 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5521 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5581 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5641 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5701 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5761 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5821 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5881 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5941 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 6001 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 6061 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 6121 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 6181 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6241 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6301 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6361 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6421 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6481 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6541 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 8041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 8101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 8161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 8221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 8761 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8821 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8881 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 8941 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 9001 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 9061 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 9121 tagt //