LOCUS Exported 5619 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CFP-FKBP (CF) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5619) AUTHORS Inoue T, Heo WD, Grimley JS, Wandless TJ, Meyer T TITLE An inducible translocation strategy to rapidly activate and inhibit small GTPase signaling pathways. JOURNAL Nat Methods. 2005 Jun . 2(6):415-8. PUBMED 15908919 REFERENCE 2 (bases 1 to 5619) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5619 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 547..556 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 553..1269 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(598..619) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(859..878) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1206..1227 /label=EGFP-C /note="EGFP, forward primer" CDS 1282..1605 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" polyA_signal 2347..2468 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2384..2403) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2438..2457 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2475..2930) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2612..2633) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2824..2843 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2957..3061 /gene="bla" /label=AmpR promoter promoter 3063..3420 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3086..3106) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3271..3406 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3333..3352 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3455..4249 /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(3509..3528) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4119..4138 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4437..4456) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4481..4528 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4857..5445 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5346..5365 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 721 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc 781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 961 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac 1021 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc 1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1261 ctgtacaaga gtgctggtgg tggagtgcag gtggaaacca tctccccagg agacgggcgc 1321 accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga 1381 aagaaatttg attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag 1441 gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa 1501 ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca 1561 catgccactc tcgtcttcga tgtggagctt ctaaaactgg aatgagctgg tggtagtgct 1621 ggtggtcctc gagctcaagc ttcgaattcg atgcaggcca tcaagtgtgt ggtggtggga 1681 gacggagctg taggtaaaac ttgcctactg atcagttaca caaccaatgc atttcctgga 1741 gaatatatcc ctactgtctt tgacaattat tctgccaatg ttatggtaga tggaaaaccg 1801 gtgaatctgg gcttatggga tacagctgga ctagaagatt atgacagatt acgcccccta 1861 tcctatccgc aaacagatgt gttcttaatt tgcttttccc ttgtgagtcc tgcatcattt 1921 gaaaatgtcc gtgcaaagtg gtatcctgag gtgcggcacc actgtcccaa cactcccatc 1981 atcctagtgg gaactaaact tgatcttagg gatgataaag acacgatcga gaaactgaag 2041 gagaagaagc tgactcccat cacctatccg cagggtctag ccatggctaa ggagattggt 2101 gctgtaaaat acctggagtg ctcggcgctc acacagcgag gcctcaagac agtgtttgac 2161 gaagcgatcc gagcagtcct ctgcccgcct cccgtgaaga agaggaagag aaaatgagga 2221 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt 2281 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 2341 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 2401 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 2461 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 2521 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 2581 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 2641 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 2701 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 2761 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 2821 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 2881 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 2941 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 3001 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 3061 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3241 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3301 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 3361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3421 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3481 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3541 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 3601 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 3661 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 3721 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 3781 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 3841 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 3901 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3961 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 4021 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 4081 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 4141 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4201 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4261 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4321 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4381 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 4441 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 4501 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 4561 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 4621 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 4681 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 4741 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 4981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5461 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 5521 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat 5581 tacggggtca ttagttcata gcccatatat ggagttccg //