LOCUS Exported 5385 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS mGFP-10-sREACh-N3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5385) AUTHORS Murakoshi H, Lee SJ, Yasuda R TITLE Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP. JOURNAL Brain Cell Biol. 2008 Aug . 36(1-4):31-42. PUBMED 18512154 REFERENCE 2 (bases 1 to 5385) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5385 /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 597..606 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 603..1121 /codon_start=1 /product="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN173 /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIE" primer_bind 1256..1277 /label=EGFP-C /note="EGFP, forward primer" CDS 1797..2048 /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH YLSYQSALSKDPNEKRDHMVLLERVTAAGITLGMDELYK" polyA_signal 2173..2294 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2210..2229) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2264..2283 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2301..2756) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2438..2459) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2650..2669 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2783..2887 /gene="bla" /label=AmpR promoter promoter 2889..3246 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2912..2932) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3097..3232 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3159..3178 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3281..4075 /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(3335..3354) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3945..3964 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4263..4282) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4307..4354 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4683..5271 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5172..5191 /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 gctagcgcca 601 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 781 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 1021 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 1081 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 1201 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg 1261 tcctgctgga gttcgtgacc gccgccggga tcactctcgg cgtcgacggt accgcgggcc 1321 cgggatccgg aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 1381 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 1441 atgccaccta cggcaagctg accctgaagc tcatctgcac caccggcaag ctgcccgtgc 1501 cctggcccac cctcgtgacc accttcggct acggcctgat gtgcttcgcc cgctaccccg 1561 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 1621 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 1681 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 1741 tcctggggca caagctggag tacaactgga acagcgtcaa cgtctatatc atggccgaca 1801 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 1861 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 1921 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg 1981 atcacatggt cctgctggag agagtgaccg ccgccgggat cactctcggc atggacgagc 2041 tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt 2101 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 2161 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 2221 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 2281 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 2341 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 2401 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 2461 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 2521 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 2581 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 2641 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 2701 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg 2761 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 2821 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 2881 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag 2941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3001 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3061 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 3121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 3181 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 3241 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 3301 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 3361 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 3421 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct 3481 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 3541 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 3601 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 3661 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 3721 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 3781 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac 3841 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 3901 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 3961 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 4021 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 4081 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 4141 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4201 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta 4261 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga 4321 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 4381 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 4441 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 4501 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt 4561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 4621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 4801 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 4861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 4921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 4981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 5281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 5341 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat //