LOCUS V044954 8482 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044954 VERSION V044954 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 874..1590 /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 1642..3972 /label="HGS(NM_004712)" /note="HGS(NM_004712)" /gene="HGS" polyA_signal 4049..4273 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4319..4747 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4761..5090 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5157..5951 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 6125..6246 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6295..6311) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6319..6335 /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(6343..6373) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6388..6409 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6697..7282) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7453..8313) /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(8314..8418) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc 901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc 1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc 1621 ggaggcggag ggagtgaatt catggggcga ggcagcggca ccttcgagcg tctcctagac 1681 aaggcgacca gccagctcct gttggagaca gattgggagt ccattttgca gatctgcgac 1741 ctgatccgcc aaggggacac acaagcaaaa tatgctgtga attccatcaa gaagaaagtc 1801 aacgacaaga acccacacgt cgccttgtat gccctggagg tcatggaatc tgtggtaaag 1861 aactgtggcc agacagttca tgatgaggtg gccaacaagc agaccatgga ggagctgaag 1921 gacctgctga agagacaagt ggaggtaaac gtccgtaaca agatcctgta cctgatccag 1981 gcctgggcgc atgccttccg gaacgagccc aagtacaagg tggtccagga cacctaccag 2041 atcatgaagg tggaggggca cgtctttcca gaattcaaag agagcgatgc catgtttgct 2101 gccgagagag ccccagactg ggtggacgct gaggaatgcc accgctgcag ggtgcagttc 2161 ggggtgatga cccgtaagca ccactgccgg gcgtgtgggc agatattctg tggaaagtgt 2221 tcttccaagt actccaccat ccccaagttt ggcatcgaga aggaggtgcg cgtgtgtgag 2281 ccctgctacg agcagctgaa caggaaagcg gagggaaagg ccacttccac cactgagctg 2341 ccccccgagt acctgaccag ccccctgtct cagcagtccc agctgccccc caagagggac 2401 gagacggccc tgcaggagga ggaggagctg cagctggccc tggcgctgtc acagtcagag 2461 gcggaggaga aggagaggct gagacagaag tccacgtaca cttcgtaccc caaggcggag 2521 cccatgccct cggcctcctc agcgcccccc gccagcagcc tgtactcttc acctgtgaac 2581 tcgtcggcgc ctctggctga ggacatcgac cctgagctcg cacggtatct caaccggaac 2641 tactgggaga agaagcagga ggaggctcgc aagagcccca cgccatctgc gcccgtgccc 2701 ctgacggagc cggctgcaca gcctggggaa gggcacgcag cccccaccaa cgtggtggag 2761 aaccccctcc cggagacaga ctctcagccc attcctccct ctggtggccc ctttagtgag 2821 ccacagttcc acaatggcga gtctgaggag agccacgagc agttcctgaa ggcgctgcag 2881 aacgccgtca ccaccttcgt gaaccgcatg aagagtaacc acatgcgggg ccgcagcatc 2941 accaatgact cggccgtgct ctcactcttc cagtccatca acggcatgca cccgcagctg 3001 ctggagctgc tcaaccagct ggacgagcgc aggctgtact atgaggggct gcaggacaag 3061 ctggcacaga tccgcgatgc ccggggggcg ctgagtgccc tgcgcgaaga gcaccgggag 3121 aagcttcgcc gggcagccga ggaggcagag cgccagcgcc agatccagct ggcccagaag 3181 ctggagataa tgcggcagaa gaagcaggag tacctggagg tgcagaggca gctggccatc 3241 cagcgcctgc aggagcagga gaaggagcgg cagatgcggc tggagcagca gaagcagacg 3301 gtccagatgc gcgcgcagat gcccgccttc cccctgccct acgcccagct ccaggccatg 3361 cccgcagccg gaggtgtgct ctaccagccc tcgggaccag ccagcttccc cagcaccttc 3421 agccctgccg gctcggtgga gggctcccca atgcacggcg tgtacatgag ccagccggcc 3481 cctgccgctg gcccctaccc cagcatgccc agcactgcgg ctgatcccag catggtgagt 3541 gcctacatgt acccagcagg ggccactggg gcgcaggcgg ccccccaggc ccaggccgga 3601 cccaccgcca gccccgctta ctcatcctac cagcctactc ccacagcggg ctaccagaac 3661 gtggcctccc aggccccaca gagcctcccg gccatctctc agcctccgca gtccagcacc 3721 atgggctaca tggggagcca gtcagtctcc atgggctacc agccttacaa catgcagaat 3781 ctcatgacca ccctcccaag ccaggatgcg tctctgccac cccagcagcc ctacatcgcg 3841 gggcagcagc ccatgtacca gcagatggca ccctctggcg gtccccccca gcagcagccc 3901 cccgtggccc agcaaccgca ggcacagggg ccgccggcac agggcagcga ggcccagctc 3961 atttcattcg actgaggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg 4021 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 4081 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 4141 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 4201 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 4261 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 4321 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 4381 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 4441 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 4501 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 4561 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 4621 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 4681 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 4741 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 4861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 4921 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 4981 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 5041 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 5101 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 5161 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 5221 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 5281 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 5341 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 5401 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 5461 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 5521 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 5581 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 5641 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 5701 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 5761 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 5821 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 5881 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 5941 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 6001 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 6061 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 6121 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 6181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 6241 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 6301 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 6361 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 6421 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 6481 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 6541 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 6601 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 6661 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 6721 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 6781 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 6841 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 6901 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6961 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 7021 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 7081 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 7141 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 7201 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 7261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 7321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 7801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 8041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 8101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 8161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 8461 agatctcccg atcccctatg gt //