LOCUS V038924 9934 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038924 VERSION V038924 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="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3692..3748 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 3749..4702 /label="OR51E1(NM_152430)" /note="OR51E1(NM_152430)" /gene="OR51E1" promoter 4722..5221 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5242..5841 /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 5855..6443 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6650..7283 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7412..7428) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7436..7452 /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(7460..7490) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7505..7526 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7814..8402) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8573..9433) /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(9434..9538) /gene="bla" /label="AmpR promoter" polyA_signal 9586..9720 /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 tacggcgtgc 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 gtacaactac 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 atctcgaggc ggcggaggga gtggaggagg 3601 gggaagcgga ggcggaggga gtggatccat gaatggcaca gaaggcccta acttctacgt 3661 gcccttctcc aatgcgacgg gtgtggtacg cgccactaac ttctccctgt tgaaacaagc 3721 aggggatgtc gaagagaatc ccgggccaat gatggtggat cccaatggca atgaatccag 3781 tgctacatac ttcatcctaa taggcctccc tggtttagaa gaggctcagt tctggttggc 3841 cttcccattg tgctccctct accttattgc tgtgctaggt aacttgacaa tcatctacat 3901 tgtgcggact gagcacagcc tgcatgagcc catgtatata tttctttgca tgctttcagg 3961 cattgacatc ctcatctcca cctcatccat gcccaaaatg ctggccatct tctggttcaa 4021 ttccactacc atccagtttg atgcttgtct gctacagatg tttgccatcc actccttatc 4081 tggcatggaa tccacagtgc tgctggccat ggcttttgac cgctatgtgg ccatctgtca 4141 cccactgcgc catgccacag tacttacgtt gcctcgtgtc accaaaattg gtgtggctgc 4201 tgtggtgcgg ggggctgcac tgatggcacc ccttcctgtc ttcatcaagc agctgccctt 4261 ctgccgctcc aatatccttt cccattccta ctgcctacac caagatgtca tgaagctggc 4321 ctgtgatgat atccgggtca atgtcgtcta tggccttatc gtcatcatct ccgccattgg 4381 cctggactca cttctcatct ccttctcata tctgcttatt cttaagactg tgttgggctt 4441 gacacgtgaa gcccaggcca aggcatttgg cacttgcgtc tctcatgtgt gtgctgtgtt 4501 catattctat gtacctttca ttggattgtc catggtgcat cgctttagca agcggcgtga 4561 ctctccgctg cccgtcatct tggccaatat ctatctgctg gttcctcctg tgctcaaccc 4621 aattgtctat ggagtgaaga caaaggagat tcgacagcgc atccttcgac ttttccatgt 4681 ggccacacac gcttcagagc cctagtctag ataattctac cgggtagggg aggcgctttt 4741 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag 4801 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt 4861 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc 4921 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca 4981 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca 5041 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct 5101 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg 5161 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc 5221 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc 5281 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg 5341 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc 5401 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg 5461 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg 5521 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg 5581 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac 5641 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc 5701 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg 5761 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg 5821 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg 5881 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 5941 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 6001 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 6061 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 6121 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 6181 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 6241 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 6301 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 6361 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 6421 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat 6481 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca 6541 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact 6601 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta 6661 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac 6721 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga 6781 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag 6841 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg 6901 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg 6961 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt 7021 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa 7081 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct 7141 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 7201 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 7261 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata 7321 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt 7381 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 8041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 8281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 8341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 8401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 9001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 9301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 9361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9541 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 9601 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 9661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 9721 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 9781 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 9841 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 9901 gtatttgtta aatatgtact acaaacttag tagt //