LOCUS Exported 5695 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CF-iSH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5695) AUTHORS Suh BC, Inoue T, Meyer T, Hille B TITLE Rapid chemically induced changes of PtdIns(4,5)P2 gate KCNQ ion channels. JOURNAL Science. 2006 Dec 1. 314(5804):1454-7. PUBMED 16990515 REFERENCE 2 (bases 1 to 5695) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5695 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" CDS 1342..1662 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" polyA_signal 2483..2604 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2520..2539) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2574..2593 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2611..3066) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2748..2769) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2960..2979 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3093..3197 /gene="bla" /label=AmpR promoter promoter 3199..3556 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3222..3242) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3407..3542 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3469..3488 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3591..4385 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3645..3664) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4255..4274 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4573..4592) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4617..4664 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4993..5581 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5482..5501 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac 1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaaga gtgctggtgg tggagtgcag gtggaaacca tctccccagg agacgggcgc 1381 accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga 1441 aagaaatttg attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag 1501 gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa 1561 ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca 1621 catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaagtgctgg tggtagtgct 1681 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtcctcgagc atccaagtac 1741 caacaagacc aggtggtgaa ggaggacagc gtagaggctg tgggcgccca gctcaaggtc 1801 taccaccagc agtaccagga caagagccgc gaatatgacc agctgtatga agaatacaca 1861 cggacctccc aggagctgca gatgaagcgc acagccatag aggccttcaa cgagaccatc 1921 aagatcttcg aagagcaggg ccagacacag gagaagtgca gcaaggagta tttggagcgc 1981 ttccggcgag agggaaatga gaaggagatg cagaggatcc tgctgaactc cgagcgactc 2041 aagtctcgca tcgcggagat acacgaaagc cgcacgaagt tggagcagga tctgcgggcg 2101 caggcctccg acaaccgtga gatcgacaag cgcatgaaca gcctcaaacc tgacctcatg 2161 cagctgcgca agatcaggga ccagtacctc gtgtggctca cccagaaagg tgcccgacag 2221 aggaagatca acgaatggct gggaatcaag aacgagactg aggaccagta ttcactgatg 2281 gaggatgagg acgccctccc ccaccacgag gagcgcacgt gagaattctg cagtcgacgg 2341 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt 2401 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 2461 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 2521 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 2581 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat 2641 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 2701 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 2761 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 2821 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 2881 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 2941 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 3001 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 3061 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 3121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 3181 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg 3241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3301 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3361 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 3421 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 3481 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 3541 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac 3601 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3661 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3721 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg 3781 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3841 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3901 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3961 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 4021 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 4081 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc 4141 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 4201 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4261 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4321 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4381 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4441 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4501 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag 4561 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc 4621 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg 4681 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 4741 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 4801 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga 4861 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 4921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 4981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 5041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 5101 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt 5161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 5221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 5281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 5341 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 5401 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 5461 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 5521 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 5581 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 5641 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat //