LOCUS V020833 5509 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020833 VERSION V020833 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 27..330 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 331..534 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" intron 672..768 /note="modified SV40 intron with splice donor and acceptor sites" /label="SV40 intron" CDS 832..855 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 859..1896 /label="Hmg20a(NM_025812)" /note="Hmg20a(NM_025812)" /gene="Hmg20a" CDS 1918..2637 /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" polyA_signal 2648..2782 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(2893..2909) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 2917..2933 /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(2941..2971) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 2986..3007 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3295..3883) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4054..4914) /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(4915..5019) /gene="bla" /label="AmpR promoter" primer_bind 5493..5509 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc 61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac 601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag 661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg 721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa 781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggactacaag 841 gacgacgatg acaagcttat ggagagcttg atggccagtt ctaccctgcc gcccctgttt 901 gcagatgaag atggctccaa ggagagcaat gatctggcta catctgggtt aacacacccc 961 gagggtccgt atggcagtgc agccacatca acaaccaacc cagagtttgt ggaggatctc 1021 tcccaaggcc agctgcttca gagtgaggct tccaatgctg tagaaggcaa tgagcaaagg 1081 cctgaagatg agcaaagaag taaacgagga ggttggtcca aagggagaaa aaggaagaaa 1141 cctcttcgag acagcaatgc acctaaatcc ccccttacag gatatgttcg attcatgaac 1201 gagcgtagag aacagcttcg agcaaagaga ccagaggttc catttccaga aatcacaagg 1261 atgttgggca atgagtggag taaactgcct cctgaggaaa agcagcgcta ccttgatgaa 1321 gcagacagag ataaggagcg ttacatgaag gaactggagc agtatcagaa gaccgaggcc 1381 tacaaggtct tcagtaggaa aacacaggac cgtcagaaag gcaaatctca taggcaagat 1441 gcagcccgac aggccactca tgatcatgag aaagaaacag aggtaaagga acgctctgtt 1501 tttgatatcc ctatatttac cgaagaattc ctgaaccaca gcaaagctcg tgaggcagag 1561 ctccgccagc ttcgtaaatc caacatggag tttgaagaga ggaacgcagc cctacagaag 1621 cacgtggaga gcatgcgcac agcagtggag aagctggagg ttgacgtgat ccaggagcgg 1681 agccgaaaca ccgtccttca gcagcacctg gagaccctgc ggcagatgtt gaccagcagc 1741 ttcgccagca tgcccttgcc tggaagtgga gaaataccaa cagtggacac cattgactca 1801 tacatgaaca gattgcacag tataatttta gctaatcccc aagacaacga aaacttcata 1861 gctacagtcc gagaggttgt gaacaggctt gatcgtggta ccggaggagg gggaagcatg 1921 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 1981 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 2041 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 2101 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 2161 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 2221 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 2281 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 2341 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 2401 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 2461 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 2521 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 2581 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtagcgg 2641 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat 2701 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat 2761 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca 2821 gggggaggtg tgggaggttt tttcggatcc tctagagtcg atctgcaggc atgctagctt 2881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 2941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 3001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 3061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 3121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 3181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 3241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 3301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 3361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 3421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 3481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 3541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 3601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 3661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 3721 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 3781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 3961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 4021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 4081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 4141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 4201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 4261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 4321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 4381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 4441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 4501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 4561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 4621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 4681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 4741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 4801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 4861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 4921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 4981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 5041 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 5101 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 5161 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 5221 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 5281 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 5341 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 5401 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 5461 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagt //