LOCUS V024056 7698 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024056 VERSION V024056 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 1660..3162 /label="Neu4(NM_001310769)" /note="Neu4(NM_001310769)" /gene="Neu4" polyA_signal 3265..3489 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3535..3963 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3977..4306 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4373..5167 /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 5341..5462 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5511..5527) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5535..5551 /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(5559..5589) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5604..5625 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5913..6498) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6669..7529) /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(7530..7634) /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 tccggactca gatctcgagg cggcggaggg 1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggagactgc tggagctccc 1681 ttctgcttcc atgtggactc cctggtacct tgctcctact ggaaggttat ggggcccacg 1741 cgtgttccca ggagaacggt gctcttccag agggaaagga cgggcctgac ctaccgtgtg 1801 cctgcgttac tctgtgtgcc tcccaggcct actctgctgg ccttcgcgga acagcgactt 1861 agccctgatg actcccatgc ccaccgcctg gtgctacgga ggggcacgct gaccaggggc 1921 tcagtgcggt ggggcactct gagtgtactg gagactgcag tactggagga gcacaggtct 1981 atgaaccctt gcccggtgct ggatgagcac tctggtacca tcttcctctt cttcattgcc 2041 gtgctgggcc acacaccgga ggccgtgcaa atcgccactg gcaagaacgc tgctcgcctc 2101 tgctgtgtga ccagctgtga cgctggcctc acctggggca gtgttcgaga tctcactgag 2161 gaagccattg gtgctgcatt gcaggactgg gccacctttg ctgtgggtcc gggccatgga 2221 gttcagctgc gctcgggtcg cctgcttgtt cctgcttaca cctatcatgt ggaccgacgg 2281 gaatgttttg gcaagatctg ctggaccagt ccccactcct tggcattcta cagtgatgat 2341 catgggatct cctggcattg tggaggcctt gtgcccaacc tacgctctgg agagtgccaa 2401 ctggctgcgg tagatggaga ctttctctac tgtaatgctc gaagccctct gggtaaccgt 2461 gtgcaggcac tgagtgctga tgaaggcacg tccttcctac caggggagct ggtgcctaca 2521 ttggcagaga cggctcgtgg ttgccagggt agcattgtgg gcttcctagc tccaccctca 2581 atcgagcctc aggatgaccg gtggacaggg agtcctagga acaccccaca ttccccatgc 2641 ttcaatctca gagtacagga gtcttcgggg gaaggtgcca gaggtcttct tgaacgttgg 2701 atgcccaggt tgcctctctg ctacccacag tcccggagcc cagagaatca tggcctagag 2761 cctgggtcag atggagataa gacatcctgg actccggaat gtcctatgtc ctctgattcc 2821 atgcttcaga gccccacatg gctactatat tcccacccag cagggcgtag agctcggctc 2881 cacatgggaa tctacctgag ccgatccccc ttggatcccc acagctggac agagccctgg 2941 gtgatctatg agggccccag tggctactct gaccttgcct ttcttgggcc tatgcctggg 3001 gcatccctgg tttttgcctg tctgtttgag agcgggacca ggacttccta tgaagacatt 3061 tctttttgct tgttctcact ggcggatgtc ctggagaatg tgcccactgg cttagagatg 3121 ctaagtctca gggataaggc tcaggggcat tgctggccct cttgaggatc cactagtcca 3181 gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg 3241 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 3301 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 3361 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 3421 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 3481 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3541 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3601 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3661 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3721 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3781 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3841 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3901 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3961 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 4141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 4261 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 4321 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 4381 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4441 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4501 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4561 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4621 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4681 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4741 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4801 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4861 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 4921 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4981 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5041 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5101 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 5161 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 5221 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 5281 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 5341 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 5401 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 5461 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 5521 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5581 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5641 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5701 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5761 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5821 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 5881 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 5941 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6001 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6061 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6121 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6181 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6241 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6301 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6361 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6421 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc 6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 7021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 7081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 7141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 7201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 7261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 7321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 7681 ctcccgatcc cctatggt //