LOCUS Exported 8912 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CMV-Brainbow-2.1 R SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8912) AUTHORS Livet J, Weissman TA, Kang H, Draft RW, Lu J, Bennis RA, Sanes JR, Lichtman JW TITLE Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. JOURNAL Nature. 2007 Nov 1. 450(7166):56-62. PUBMED 17972876 REFERENCE 2 (bases 1 to 8912) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8912 /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" protein_bind 617..650 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 673..1389 /codon_start=1 /product="humanized Renilla green fluorescent protein" /label=hrGFP /note="mammalian codon-optimized" /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE DGGLVEIRSDINLIEGMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI AQLTSLGKPLGSLHEWV" CDS 1441..1461 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter complement(1501..1519) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 1545..1656 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 1789..1910 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" CDS complement(2033..2752) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" regulatory 2749..2758 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2766..2799) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 2820..2853 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 2869..2878 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2873..2882 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(4326..4344) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 4370..4481 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 4614..4735 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" CDS complement(4850..5569) /codon_start=1 /product="enhanced monomeric variant of CFP (Rizzo et al., 2004)" /label=mCerulean /note=" mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" regulatory 5566..5575 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(5698..5731) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 5746..5779 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" rep_origin complement(5828..6283) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5965..5986) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6177..6196 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6310..6414 /gene="bla" /label=AmpR promoter promoter 6416..6773 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6439..6459) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6624..6759 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6686..6705 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6808..7602 /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(6862..6881) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7472..7491 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7790..7809) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7834..7881 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8210..8798 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8699..8718 /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 ccggactcag atcgagataa cttcgtatag catacattat acgaagttat accggatagg 661 gagacccgca ccatggtgag caagcagatc ctgaagaaca ccggcctgca ggagatcatg 721 agcttcaagg tgaacctgga gggcgtggtg aacaaccacg tgttcaccat ggagggctgc 781 ggcaagggca acatcctgtt cggcaaccag ctggtgcaga tccgcgtgac caagggcgcc 841 cccctgccct tcgccttcga catcctgagc cccgccttcc agtacggcaa ccgcaccttc 901 accaagtacc ccgaggacat cagcgacttc ttcatccaga gcttccccgc cggcttcgtg 961 tacgagcgca ccctgcgcta cgaggacggc ggcctggtgg agatccgcag cgacatcaac 1021 ctgatcgagg ggatgttcgt gtaccgcgtg gagtacaagg gccgcaactt ccccaacgac 1081 ggccccgtga tgaagaagac catcaccggc ctgcagccca gcttcgaggt ggtgtacatg 1141 aacgacggcg tgctggtggg ccaggtgatc ctggtgtacc gcctgaacag cggcaagttc 1201 tacagctgcc acatgcgcac cctgatgaag agcaagggcg tggtgaagga cttccccgag 1261 taccacttca tccagcaccg cctggagaag acctacgtgg aggacggcgg cttcgtagag 1321 cagcacgaga ccgccatcgc ccagctgacc agcctgggca agcccctggg cagcctgcac 1381 gagtgggtgg agctctccgg actcagatct cgaccgcggg cccgcgaccc tcccgtgggc 1441 cccaagaaga agcgcaaggt gggcgcctaa gatccaccgg atctagataa tagggtacca 1501 cctttagtga gggttaattg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg 1561 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactct 1621 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 1681 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 1741 gcatgctggg gcgccggcgc gttaaggcgt aaattgtaag cgttaattta agatacattg 1801 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 1861 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 1921 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt 1981 aaaacctcta caaatgtggt atggctgatt atgatctaga gtcgcggccg ctttacttgt 2041 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt 2101 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt 2161 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct 2221 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct 2281 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca 2341 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt 2401 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg 2461 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt 2521 ggtcggggta gcgggcgaag cactgcaggc cgtagccgaa ggtggtcacg agggtgggcc 2581 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg 2641 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct 2701 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga 2761 ccggtataac ttcgtataat gtatgctata cgaagttatg tcgactgcag aatttcgaga 2821 taacttcgta tagcatacat tatacgaagt tataccggct agtgattcgc caccatggtg 2881 gcctcctccg aggacgtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc 2941 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 3001 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg 3061 tccccccagt tccagtacgg ctccaaggcg tacgtgaagc accccgccga catccccgac 3121 tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 3181 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag 3241 gtgaagttcc gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg 3301 ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc 3361 caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac 3421 atggccaaga agcccgtgca gctgcccggc tactactacg tggacaccaa gctggacatc 3481 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac 3541 cacctgttcc tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc 3601 tccgaggacg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac 3661 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc 3721 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc 3781 cagttccagt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgactacaag 3841 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc 3901 gtggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag 3961 ttccgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg 4021 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag 4081 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc 4141 aagaagcccg tgcagctgcc cggctactac tacgtggaca ccaagctgga catcacctcc 4201 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg 4261 ttcctgtagg aattggccgc gactctagga ttgatctaga gtcgcggcta gataataggg 4321 taccaccttt agtgagggtt aattgctaga gctcgctgat cagcctcgac tgtgccttct 4381 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 4441 actctcactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 4501 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 4561 agcaggcatg ctggggcgcc ggcgcgttaa ggcgtaaatt gtaagcgtta atttaagata 4621 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 4681 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa 4741 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag 4801 caagtaaaac ctctacaaat gtggtatggc tgattatgat ctagccgctt tacttgtaca 4861 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat 4921 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg 4981 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca 5041 cgctgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct 5101 tgtcggcggt gatatagacg ttgtcgctga tggcgttgta ctccagcttg tgccccagga 5161 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc 5221 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc 5281 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt 5341 cggggtagcg ggcgaagcac tgcacgcccc aggtcagggt ggtcacgagg gtgggccagg 5401 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat 5461 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga 5521 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg 5581 gtggatcccg ggcccgcggt accgtcgact gcagctcctg atcggcctct tcattcttct 5641 caaccggttt agttcttctg atgcagcaca gcatggtggc tgtgaattaa ttccggtata 5701 acttcgtata atgtatgcta tacgaagtta tctcgaatcg cgcgcgaagt tcctattctc 5761 tagaaagtat aggaacttct cgaagcttga gctcgagatc tgagtccggt agcgctagtt 5821 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc 5881 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag 5941 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg 6001 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca 6061 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa 6121 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg 6181 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta 6241 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg 6301 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 6361 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag 6421 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 6481 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 6541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 6601 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 6661 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6721 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat 6781 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 6841 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 6901 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 6961 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc 7021 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 7081 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 7141 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 7201 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 7261 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 7321 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 7381 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 7441 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 7501 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 7561 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 7621 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 7681 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 7741 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga 7801 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 7861 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg 7921 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc 7981 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg 8041 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta 8101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 8161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 8221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 8281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 8341 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 8401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 8461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 8521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 8581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 8641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 8701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 8761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 8821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 8881 attctgtgga taaccgtatt accgccatgc at //