LOCUS Exported 8239 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAG-Brainbow3.0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8239) AUTHORS Cai D, Cohen KB, Luo T, Lichtman JW, Sanes JR TITLE Improved tools for the Brainbow toolbox. JOURNAL Nat Methods. 2013 May 5;10(6):540-7. doi: 10.1038/nmeth.2450. Epub 2013 May 5. PUBMED 23817127 REFERENCE 2 (bases 1 to 8239) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8239 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 662..1678 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1602..1624 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1686..1705 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 1745..1769 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind 1818..1851 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 1860..1893 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." regulatory 1902..1911 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1908..2615 /codon_start=1 /product="photostable monomeric orange derivative of DsRed fluorescent protein (Shaner et al., 2008)" /label=mOrange2 /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" polyA_signal 2797..2918 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 2931..2964 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 2973..2982 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2979..3695 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2979..3695 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3024..3045) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3285..3304) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3632..3653 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3877..3998 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 4015..4048 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." regulatory 4057..4066 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4063..4758 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" polyA_signal 4940..5061 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 5074..5107 /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)" protein_bind complement(5146..5170) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(5261..5280) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 5326..5381 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(5380..5399) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5742..5758) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5742..5758) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5755..5777) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5766..5782 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5790..5820) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5835..5856 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5915..6110 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5961..6096 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6023..6042 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 6116..6250 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6318..6335) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6489..7077) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6569..6588) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7248..8108) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7871..7890 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8109..8213) /gene="bla" /label=AmpR promoter ORIGIN 1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccagggggg gggggggggg gggggggggg 541 gggggggggg gggggggggg gggcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga 841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg 901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg 961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg 1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt 1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca 1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg 1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc 1261 ggggccgcct cgggccgggg ggggctcggg ggaggggcgc ggcggccccc ggagcgccgg 1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc 1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac 1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga 1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc 1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt 1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct 1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgagga 1741 tatcacaagt ttgtacaaaa aagcaggctt taaaggaacg cacgtaacta taacggtcct 1801 aaggtagcga acctaggata acttcgtata gcatacatta tacgaagtta tcggcgcgca 1861 taacttcgta taggatactt tatacgaagt tatcaagcgc tgccaccatg gtgagcaagg 1921 gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg cgcatggagg 1981 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg 2041 gctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca 2101 tcctgtcccc tcatttcacc tacggctcca aggcctacgt gaagcacccc gccgacatcc 2161 ccgactactt caagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaactacg 2221 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct 2281 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtgatg cagaagaaga 2341 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggtgcc ctgaagggca 2401 agatcaagat gaggctgaag ctgaaggacg gcggccacta cacctccgag gtcaagacca 2461 cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac atcaagttgg 2521 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc 2581 gccactccac cggcggcatg gacgagctgt acaagaagct gaaccctcct gatgagagtg 2641 gccccggctg catgagctgc tgtgtgctct cctgagcggc cgcgactcta gatcataatc 2701 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 2761 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 2821 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 2881 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtgctagc ataacttcgt 2941 atagcataca ttatacgaag ttatctaacg ttgccaccat ggtgagcaag ggcgaggagc 3001 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 3061 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 3121 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg 3181 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 3241 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 3301 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 3361 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 3421 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 3481 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 3541 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc 3601 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 3661 ccgggatcac tctcggcatg gacgagctgt acaagaagct gaaccctcct gatgagagtg 3721 gccccggctg catgagctgc tgtgtgctct cctgagcggc cgcgactcta gatcataatc 3781 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 3841 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 3901 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 3961 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtgtttaa acatataact 4021 tcgtatagga tactttatac gaagttatcg cgtacggcca ccatggtgag cgagctgatt 4081 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag 4141 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggcg 4201 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc 4261 agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttcccc 4321 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc 4381 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac 4441 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc ctccaccgag 4501 accctgtacc ccgctgacgg cggcctggaa ggcagagccg acatggccct gaagctcgtg 4561 ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa acccgctaag 4621 aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat caaggaggcc 4681 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct 4741 agcaaactgg ggcacagaaa gctgaaccct cctgatgaga gtggccccgg ctgcatgagc 4801 tgctgtgtgc tctcctgagc ggccgcgact ctagatcata atcagccata ccacatttgt 4861 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 4921 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 4981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5041 caaactcatc aatgtatctt aaggcgtact agtgaagttc ctattctcta gaaagtatag 5101 gaacttccgt aactataacg gtcctaaggt agcgaatgca tctagaccca gctttcttgt 5161 acaaagtggt gattcgcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg 5221 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa 5281 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta 5341 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga 5401 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc 5461 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc 5521 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta 5581 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac 5641 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat 5701 ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg 5761 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5821 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5881 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag 5941 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 6001 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 6061 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt 6121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6241 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 6301 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 6361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 6421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 6481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 6541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 6601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 6661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 6721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 6781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 6841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 6901 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 6961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 7021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 7081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 7141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 7201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 7261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 7321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 7381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 7441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 7501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 7561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 7621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 7681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 7741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 7801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 7861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 7921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 7981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 8041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 8101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 8161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 8221 ccccgaaaag tgccacctg //