LOCUS V020626 7338 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020626 VERSION V020626 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..2850 /label="P2RX3(NM_002559)" /note="P2RX3(NM_002559)" /gene="P2RX3" polyA_signal 2905..3129 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3175..3603 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3617..3946 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4013..4807 /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 4981..5102 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5151..5167) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5175..5191 /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(5199..5229) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5244..5265 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5553..6138) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6309..7169) /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(7170..7274) /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 tgaactgcat atccgacttc 1681 ttcacctatg agaccaccaa gtcggtggtt gtgaagagct ggaccatcgg gatcatcaac 1741 cgagtagttc agcttctgat catctcctac tttgtagggt gggttttctt gcacgagaag 1801 gcttaccagg tacgggacac agccattgag tcctcggtgg taaccaaggt gaagggctcc 1861 ggactctacg ccaacagagt catggatgtg tctgattacg tgacgccacc tcagggcacc 1921 tcggtctttg tcatcatcac caagatgatt gttactgaaa atcagatgca aggattctgc 1981 ccagagagtg aggagaaata ccgctgtgta tcagacagcc agtgcgggcc tgagcgcttg 2041 ccaggtgggg ggatcctcac tggccgctgc gtgaactaca gctctgtgct ccggacctgt 2101 gagatccagg gctggtgccc cacggaggtg gacacagtgg aaacgcccat catgatggaa 2161 gctgagaact tcactatttt catcaagaac agcatccgtt tccccctctt caactttgag 2221 aagggaaacc tccttcccaa cctgacagcc agggacatga agacctgccg cttccacccg 2281 gacaaggacc ctttctgccc catcttgcgg gtaggggacg tggtcaagtt tgcggggcag 2341 gattttgcca aactggcgcg cacgggggga gttctgggca ttaagatcgg ctgggtgtgc 2401 gacttggaca aggcctggga ccagtgcatc cccaaatact ccttcacccg gctcgacagc 2461 gtttctgaga aaagcagcgt gtccccaggc tacaacttca ggtttgccaa gtactacaaa 2521 atggaaaatg gcagtgagta ccgcaccctc ctgaaggctt ttggcatccg cttcgacgtg 2581 ctggtatacg ggaatgctgg caagttcaac atcatcccca ccatcatcag ctctgtggcg 2641 gcctttactt ctgtgggagt gggaactgtt ctctgtgaca tcatcctgct caacttcctc 2701 aagggggccg accagtacaa agccaagaag tttgaggagg tgaatgagac tacgctgaaa 2761 atcgcggctt tgaccaaccc agtgtacccc agcgaccaga ccacagcgga gaagcagtcc 2821 accgattcgg gggccttctc cataggccac tagggcggcc gctcgagtct agagggcccg 2881 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2941 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 3001 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 3061 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 3121 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3241 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3361 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3481 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3541 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3601 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3721 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3781 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3841 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3901 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3961 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 4021 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4081 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4141 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4201 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4261 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4321 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4381 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4441 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4501 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 4561 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4621 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4681 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4741 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4801 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4861 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4921 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 5041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 5101 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 5161 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5221 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 5521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 5581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6061 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc 6121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 7321 ctcccgatcc cctatggt //