LOCUS V041581 10250 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041581 VERSION V041581 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 2829..2849 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" CDS 2850..4253 /label="Gsdmc(NM_031378)" /note="Gsdmc(NM_031378)" /gene="Gsdmc" CDS 4290..5009 /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" promoter 5038..5537 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5558..6157 /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 6171..6759 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6966..7599 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7728..7744) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7752..7768 /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(7776..7806) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7821..7842 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8130..8718) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8889..9749) /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(9750..9854) /gene="bla" /label="AmpR promoter" polyA_signal 9902..10036 /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 ccaccatgcc caagaagaag aggaaggtga tgtcctacac atttgactgg cttagcaagg 2881 atgtggttaa aaagcttcaa ggaagagatc tgaggcctgt caaatgcctg tcagatgcaa 2941 ccaaattctg cctgtttaac atacttcagg aaacttcttc ccggttggct ttgaaaactg 3001 aatatattcc agttggattt acccttctgc atctcttgga gcccaatatt cctgttccag 3061 agcctgaagt gtcagctccc atccccctta aacacaccat atcccagaaa ctgaaggctg 3121 acctggatgt cgaaaccatt gcaggaggag aagcagggtt tgttaagtcc tgtggatatg 3181 acatcgaggt tcagagtaag agcatcccaa atccaaaact ggaaagtctt caaaacagaa 3241 aactgttgga tcagttgccc acatttatga agacttgctg gaaggatgga aagaacctgt 3301 atgtggtgac agaggcctat gaagtgacca aggatactgt gctggaaggc acaagcaata 3361 gcaaatttgc aatcaaaggg atcatcaacc agcttgtcaa ggttggaggc agtggacagt 3421 ggcaaacaga gaaaacagat tcaataccta tccaaaaggg ttcagtgttg gcctacaaga 3481 agcagcaatt ggtcattgag gataacactt gtgttattct cacttctgct aacactaaga 3541 agaagatgac ctttccaatg agatttgtgg gaatgtctgg acatctaaga tatcaggatc 3601 tagtcattga gacaggctcg tggataaatg acatcgatcc aattggaaca attaaggagc 3661 ccacccatct agatttcatg tgcctacaaa atgaagtttc tgaacaaaca cggctactgg 3721 ctgaactctc aaaggatgtt caagaagttg tgttctccag tttcctgcac atgctctgtg 3781 acagggatgt tctgtatgac ctgatgaaaa tgctggagtt gaatcagttg gggcatatgg 3841 atggcccagg tggcaaaatc ctggatgagc ttcgaaagga ctcgagcctt tcatggatta 3901 acctaaagga cctcattctt tatctccttc aagccctgat ggtgctgagt gatacccaac 3961 tctgtctgct ggccctgtct gtggagatgc ggctcctccc gcaccaagtg gagctggtaa 4021 agagcatctt acagccaaac ttcaagtatc cctggaacat tcccttcaca ctccagcctc 4081 agcttcttgc cccactccag ggtgagggct tggccattac ttatgaattg ctggaggagt 4141 gtggtctgaa gatggagctg aataatccca ggtcaacttg ggatttggaa gccaagatgc 4201 ccctgtctgc tctctatgga agcctgtcat ttttacagca actttcagaa gccggtggag 4261 gcggatcagg aggcggtgga tcgggcccga tggtgagcaa gggcgaggag ctgttcaccg 4321 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 4381 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 4441 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 4501 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 4561 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 4621 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 4681 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 4741 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 4801 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 4861 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 4921 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 4981 ctctcggcat ggacgagctg tacaagtagg gatcccgcga ctctagataa ttctaccggg 5041 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca 5101 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc 5161 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga 5221 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc 5281 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg 5341 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg 5401 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg 5461 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct 5521 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt 5581 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc 5641 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga 5701 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga 5761 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt 5821 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca 5881 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt 5941 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt 6001 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct 6061 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc 6121 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc 6181 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 6241 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 6301 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 6361 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 6421 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 6481 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 6541 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg 6601 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 6661 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc 6721 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg 6781 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg 6841 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc 6901 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag 6961 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta 7021 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata 7081 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga 7141 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga 7201 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc 7261 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact 7321 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct 7381 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga 7441 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 7501 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 7561 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta 7621 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt 7681 gctagcgttt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 7741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 7801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 7861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 7921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 7981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 8041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 8101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 8161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 8221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 8281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 8341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 8401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 8461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 8521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 8581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 8641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 8701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 8761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 8821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 8881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 8941 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 9001 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 9061 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 9121 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 9181 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 9241 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 9301 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 9361 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 9421 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 9481 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 9541 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 9601 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 9661 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 9721 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 9781 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 9841 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 9901 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 9961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 10021 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac 10081 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc 10141 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt 10201 agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt //