LOCUS V025447 7045 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025447 VERSION V025447 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 905..1855 /label="Tacstd2(NM_020047)" /note="Tacstd2(NM_020047)" /gene="Tacstd2" CDS 1895..2614 /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 2645..2869 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2915..3343 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3357..3686 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3753..4547 /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 4721..4842 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4891..4907) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4915..4931 /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(4939..4969) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4984..5005 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5293..5878) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6049..6909) /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(6910..7014) /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 caccatggcg aggggcttgg atctagcacc gctgctactg ctactgctgg cgatggcgac 961 ccgcttttgc acggctcaga gcaactgtac atgccccacc aacaagatga cggtctgcga 1021 cacaaatggc ccaggcgggg tctgccaatg tcgggcaatg ggctcacagg tattggtcga 1081 ctgctccacg ctaacttcca agtgcctgct gctcaaggcg cgcatgagcg cccggaagag 1141 cggccgcagc ctggtgatgc cgagcgagca cgcgatactg gacaacgatg gcctctacga 1201 cccggagtgt gacgacaagg gccgcttcaa ggcgcgccag tgcaaccaga cctcggtgtg 1261 ctggtgcgta aactcggtgg gcgtgcgccg cacggacaag ggagaccaaa gcctgcgctg 1321 cgacgaagtg gtgcgaaccc accacatcct cattgagttg cgccaccgcc cgaccgaccg 1381 agccttcaac cactctgacc tagactccga gctgcggcgg ctcttccaag aacgctacaa 1441 gctgcacccc agcttcctat ccgcggtaca ctatgaggag cccaccattc agatagagct 1501 tcggcagaac gcgtcgcaga agggcttgag agacgtggac atcgctgatg ccgcctacta 1561 cttcgaaagg gacattaaag gcgagtcact gttcatgggc cgccgcggcc tggacgtgca 1621 ggtgcgtggg gaacccctgc atgtggagcg gacgctcatc tactacctgg acgagaagcc 1681 cccccagttc tccatgaagc gcctcaccgc cggcgtcatt gccgtcatcg ctgtcgtctc 1741 ggtagcggta gtggctggtg tggtggtctt ggtggtcacc aaacggagga agtcgggcaa 1801 atacaaaaag gtggagctta aggagctggg ggagatgaga agcgaaccta gcttgtactc 1861 gagtggcggt ggaggaagcg gcggaggtgg atctatggtg agcaagggcg aggagctgtt 1921 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1981 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 2041 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 2101 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 2161 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 2221 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 2281 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 2341 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 2401 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 2461 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 2521 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 2581 gatcactctc ggcatggacg agctgtacaa gtagggcccg tttaaacccg ctgatcagcc 2641 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2701 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2761 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2821 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2881 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2941 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3001 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3061 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3121 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3241 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3361 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3421 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3481 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3541 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3601 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3661 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3721 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3781 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3841 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3901 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3961 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4021 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4081 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4141 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4201 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4261 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4321 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4381 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4441 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4501 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4561 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4621 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4681 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4741 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4801 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4861 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5821 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5881 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5941 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6001 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6061 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6121 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6181 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6241 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6301 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6361 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6421 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6481 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6541 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6601 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6661 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6721 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6781 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6841 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6901 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6961 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7021 tccccgaaaa gtgccacctg acgtc //