LOCUS Exported 4683 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses GFP1-9 in zebrafish. ACCESSION . VERSION . KEYWORDS pCS2-GFP1-9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4683) AUTHORS Zhang Q, Schepis A, Huang H, Yang J, Ma W, Torra J, Zhang SQ, Yang L, Wu H, Nonell S, Dong Z, Kornberg TB, Coughlin SR, Shu X TITLE Designing a green fluorogenic protease reporter by flipping a beta strand of GFP for imaging apoptosis in animals. JOURNAL J Am Chem Soc. 2019 Mar 1. doi: 10.1021/jacs.8b13042. PUBMED 30821975 REFERENCE 2 (bases 1 to 4683) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4683 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" promoter complement(710..727) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 732..866 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(756..775) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 810..829 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(979..999) /label=T3 /note="T3 promoter, forward primer" promoter complement(979..997) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1018..1034) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1018..1034) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1031..1053) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1042..1058 /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(1066..1096) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1111..1132 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1249..1266) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1420..2008) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1500..1519) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2179..3039) /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 2802..2821 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3040..3144) /gene="bla" /label=AmpR promoter rep_origin complement(3170..3625) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3307..3328) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3519..3538 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3699..4683 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atccgccacc atgcgcaaag gcgaagaact gtttaccggc 121 attgtgccga ttctggtgga actggatggc gatgtgaacg gccataaatt ttttgtgcgc 181 ggcgaaggcg aaggcgatgc gaccattggc aaactgagcc tgaaatttat ttgcaccacc 241 ggcaaactgc cggtgccgtg gccgaccctg gtgaccaccc tgacctatgg cgtgcagtgc 301 tttagccgct atccggatca tatgaaacgc catgattttt ttaaaagcgc gatgccggaa 361 ggctatgtgc aggaacgcac catttatttt aaagatgatg gcacctataa aacccgcgcg 421 gaagtgaaat ttgaaggcga taccctggtg aaccgcattg aactgaaagg cattgatttt 481 aaagaagatg gcaacattct gggccataaa ctggaatata actttaacag ccataaagtg 541 tatattaccg cggataaaca gaacaacggc attaaagcga actttaccat tcgccataac 601 gtggaagatg gcagcgtgca gctggcggat cattatcagc agaacacccc gattggcgat 661 ggcccggttc ttcttcctta agaattcaag gcctctcgag cctctagaac tatagtgagt 721 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta 781 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 841 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 901 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc 961 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc 1021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 1081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 1141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 1201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 1261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 1321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 1381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 1441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 1501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 1561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 1621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 1681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 1741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 1801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 1861 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 1921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 1981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 2041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 2101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 2161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 2221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 2281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 2341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 2401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 2461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 2521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 2581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 2641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 2701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 2761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 2821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 2881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 2941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 3001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 3061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 3121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc 3181 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 3241 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 3301 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 3361 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 3421 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 3481 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 3541 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 3601 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa 3661 gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccatagccaa ttcaatatgg 3721 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg 3781 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg 3841 cggacttggc accatgccaa ttcaatatgg cggacttggc actgtgccaa ctggggaggg 3901 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat 3961 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata 4021 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt 4081 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc 4141 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc 4201 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc 4261 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc 4321 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg 4381 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg 4441 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaaggacgcc 4501 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc 4561 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg 4621 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg 4681 aac //