LOCUS V029868 10090 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029868 VERSION V029868 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 2815..4008 /label="B4GALT1(NM_001497)" /note="B4GALT1(NM_001497)" /gene="B4GALT1" CDS 4045..4764 /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" misc_feature 4793..5379 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5399..5998 /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 6012..6600 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6807..7440 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7568..7584) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7592..7608 /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(7616..7646) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7661..7682 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7970..8558) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8729..9589) /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(9590..9694) /gene="bla" /label="AmpR promoter" polyA_signal 9742..9876 /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 tctatttccg gtgaattcgc caccatgagg 2821 cttcgggagc cgctcctgag cggcagcgcc gcgatgccag gcgcgtccct acagcgggcc 2881 tgccgcctgc tcgtggccgt ctgcgctctg caccttggcg tcaccctcgt ttactacctg 2941 gctggccgcg acctgagccg cctgccccaa ctggtcggag tctccacacc gctgcagggc 3001 ggctcgaaca gtgccgccgc catcgggcag tcctccgggg agctccggac cggaggggcc 3061 cggccgccgc ctcctctagg cgcctcctcc cagccgcgcc cgggtggcga ctccagccca 3121 gtcgtggatt ctggccctgg ccccgctagc aacttgacct cggtcccagt gccccacacc 3181 accgcactgt cgctgcccgc ctgccctgag gagtccccgc tgcttgtggg ccccatgctg 3241 attgagttta acatgcctgt ggacctggag ctcgtggcaa agcagaaccc aaatgtgaag 3301 atgggcggcc gctatgcccc cagggactgc gtctctcctc acaaggtggc catcatcatt 3361 ccattccgca accggcagga gcacctcaag tactggctat attatttgca cccagtcctg 3421 cagcgccagc agctggacta tggcatctat gttatcaacc aggcgggaga cactatattc 3481 aatcgtgcta agctcctcaa tgttggcttt caagaagcct tgaaggacta tgactacacc 3541 tgctttgtgt ttagtgacgt ggacctcatt ccaatgaatg accataatgc gtacaggtgt 3601 ttttcacagc cacggcacat ttccgttgca atggataagt ttggattcag cctaccttat 3661 gttcagtatt ttggaggtgt ctctgctcta agtaaacaac agtttctaac catcaatgga 3721 tttcctaata attattgggg ctggggagga gaagatgatg acatttttaa cagattagtt 3781 tttagaggca tgtctatatc tcgcccaaat gctgtggtcg ggaggtgtcg catgatccgc 3841 cactcaagag acaagaaaaa tgaacccaat cctcagaggt ttgaccgaat tgcacacaca 3901 aaggagacaa tgctctctga tggtttgaac tcactcacct accaggtgct ggatgtacag 3961 agatacccat tgtataccca aatcacagtg gacatcggga caccgagcct cgagggcggt 4021 ggaggaagcg gcggaggtgg atctatggtg agcaagggcg aggagctgtt caccggggtg 4081 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 4141 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 4201 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc 4261 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 4321 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 4381 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 4441 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat 4501 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 4561 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 4621 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc 4681 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 4741 ggcatggacg agctgtacaa gtagactagt tctagagcgg ccgcggatcc cgcccctctc 4801 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 4861 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 4921 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 4981 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 5041 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 5101 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 5161 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 5221 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct 5281 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 5341 ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac gaccttccat 5401 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 5461 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 5521 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 5581 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 5641 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 5701 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 5761 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 5821 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 5881 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 5941 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac 6001 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 6061 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 6121 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 6181 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 6241 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 6301 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 6361 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 6421 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 6481 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 6541 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 6601 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac 6661 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt 6721 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6781 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga 6841 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac 6901 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt 6961 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt 7021 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg 7081 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata 7141 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg 7201 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg 7261 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 7321 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 7381 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 7441 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 7501 agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag agcttggcgt 7561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7621 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7681 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7741 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 8041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 8101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 8161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 8221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 8281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 8341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 8401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 8461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 8761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 9001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 9061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 9121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 9181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 9241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 9301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 9361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 9421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9601 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9661 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9721 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9841 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9901 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9961 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 10021 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 10081 acttagtagt //