LOCUS V042585 7805 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042585 VERSION V042585 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 935..1651 /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 1688..3295 /label="Egr1(NM_012551)" /note="Egr1(NM_012551)" /gene="Egr1" CDS 3302..3367 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3405..3629 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3675..4103 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4117..4446 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4513..5307 /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 5481..5602 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5651..5667) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5675..5691 /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(5699..5729) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5744..5765 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6053..6638) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6809..7669) /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(7670..7774) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccatggtg agcaagggcg aggagctgtt 961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1141 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 1381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 1621 gatcactctc ggcatggacg agctgtacaa ggaattcggt ggaggcggat caggaggcgg 1681 tggatctatg gcagcggcca aggccgagat gcaattgatg tctccgctgc agatctctga 1741 cccgttcggc tcctttcctc actcacccac catggacaac taccccaaac tggaggagat 1801 gatgctgctg agcaacgggg ctccccagtt cctcggtgct gccggaaccc cagagggcag 1861 cggcggcaat aacagcagca gcagcagcag cagcagcagc gggggcggtg gtgggggcgg 1921 cagcaacagc ggcagcagcg ctttcaatcc tcaaggggag ccgagcgaac aaccctacga 1981 gcacctgacc acagagtcct tttctgacat cgctctgaat aacgagaagg cgctggtgga 2041 gacaagttat cccagccaaa ctacccggtt gcctcccatc acctatactg gccgcttctc 2101 cctggagcct gcacccaaca gtggcaacac tttgtggcct gaaccccttt tcagcctagt 2161 cagtggcctt gtgagcatga ccaaccctcc aacctcttca tcctcagcgc cttctccagc 2221 tgcttcatcg tcttcctctg cctcccagag cccacccctg agctgtgccg tgccgtccaa 2281 cgacagcagt cccatttact cagctgcacc cacctttcct actcccaaca ctgacatttt 2341 tcctgagccc caaagccagg cctttcctgg ctctgcaggc acagccttgc agtacccgcc 2401 tcctgcctac cctgccacca agggtggttt ccaggttccc atgatccctg actatctgtt 2461 tccacaacaa cagggagacc tgagcctggg caccccagac cagaagccct tccagggtct 2521 ggagaaccgt acccagcagc cttcgctcac tccactatcc actatcaaag ccttcgccac 2581 tcagtcgggc tcccaggact taaaggctct taataacacc taccagtccc aactcatcaa 2641 acccagccgc atgcgcaagt accccaaccg gcccagcaag acaccccccc atgaacgccc 2701 gtatgcttgc cctgttgagt cctgcgatcg ccgcttttct cgctcggatg agcttacacg 2761 ccacatccgc atccatacag gccagaagcc cttccagtgt cgaatctgca tgcgtaattt 2821 cagtcgtagt gaccacctta ccacccacat ccgcacccac acaggcgaga agccttttgc 2881 ctgtgacatt tgtgggagaa agtttgccag gagtgatgaa cgcaagaggc ataccaaaat 2941 ccacttaaga cagaaggaca agaaagcaga caaaagtgtc gtggcctcct cagctgcctc 3001 ttccctctct tcctacccat ccccagtggc tacctcctac ccatcccccg ccaccacctc 3061 atttccatcc ccagtgccca cctcttactc ctctccgggc tcctctacct acccgtctcc 3121 tgcacacagt ggcttcccat cgccctcggt ggccaccacc tatgcctccg tcccacctgc 3181 tttccctgcc caggtcagca ccttccagtc tgcaggggtc agcaactcct tcagcacctc 3241 aacgggtctt tcagacatga cagcaacctt ttctcctagg acaattgaaa tttgcctcga 3301 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 3361 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg 3421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 3481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 3541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 3601 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 3661 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 4021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 4081 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 4141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 4201 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4261 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 4321 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 4381 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 4441 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 4501 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 4561 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 4621 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 4681 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 4741 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 4801 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4861 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4921 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4981 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 5041 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 5101 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 5161 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 5221 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 5281 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 5341 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5401 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5461 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 5521 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5581 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 5641 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 5701 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 5761 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 5821 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5881 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5941 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 6001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 6061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt 6601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 7201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 7261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 7321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 7381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7801 acgtc //