LOCUS       sequence-237431-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP1-9 in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Am Chem 
            Soc. 2019 Mar 1. doi: 10.1021/jacs.8b13042."
COMMENT     Sequence Label: pCS2-GFP1-9
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     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(join(1066..1072,1073..1090,1091..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(join(2179..2970,2971..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
//
