LOCUS       sequence-363970-        8298 bp    DNA     circular SYN 15-APR-2025
DEFINITION  cyclofen-inducible apoptosis activation and imaging (mCherry) in 
            zebrafish permanent transgenic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8298)
  AUTHORS   Feng Z, Zhang W, Xu J, Gauron C, Ducos B, Vriz S, Volovitch M, 
            Jullien L, Weiss S, Bensimon D
  TITLE     Optical control and study of biological processes at the single-cell
            level in a live organism.
  JOURNAL   Rep Prog Phys. 2013 Jul;76(7):072601. doi: 
            10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14.
  PUBMED    23764902
REFERENCE   2  (bases 1 to 8298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Rep Prog 
            Phys."; date: "2013-07"; volume: "76(7):072601. doi"; pages: " 
            10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14"
COMMENT     Sequence Label: pT22iuasvApoWChe(#488)
FEATURES             Location/Qualifiers
     source          1..8298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(150..170)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(150..168)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    429..550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(570..1277,1278..1280,1281..1283))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(609..630)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     958..977
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1217..1238
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2044..2055)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2397..2438
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2934..3641
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             3654..3710
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5094..5114
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6087..6208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6549..7137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6629..6648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7308..8099,8100..8168))
                     /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     7931..7950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8169..8273)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccc gatactccag cttttgttcc ctttagtgag ggttaattgc gcgttacaca
      181 gaggtgtaaa gtacttgagt aattttactt gattactgta cttaagtatt atttttgggg
      241 atttttactt tacttgagta caattaaaaa tcaatacttt tacttttact taattacatt
      301 tttttagaaa aaaaagtact ttttactcct tacaatttta tttacagtca aaaagtactt
      361 attttttgga gatcacttcc tcatagggat aacagggtaa tggccggcct tttcaggacc
      421 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      481 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      541 taaacaagtt gcgtgcgcat ttatttaagg tacagctcgt ccatgccgag agtgatcccg
      601 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
      661 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
      721 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
      781 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
      841 ctgatgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      901 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      961 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     1021 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     1081 ccccaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     1141 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     1201 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     1261 agctcctcgc ccttgctcac catgtcggct ggcgaccggt ggatcatccc ccgcaagtct
     1321 gactgagtgg aagtttcagt tcaatggaag ggagaaaaga gccagccttt atatatgtca
     1381 tgtattgctg ctgtctgcat gcgatgaacc tagagcttac tttttctgtc aagtaaggca
     1441 aaagcagaga acctttgtct tgcattgtgt ttgatgttgt gctgtgacag tcatttctta
     1501 gttcatgctg gtacttttgt tccatagctt tacccaaaga gttatccagc attcccagtt
     1561 attccaaaaa ctcaatgtca caaattttcc agtccattgc ttagcagcgt tgaccccgga
     1621 gaccctgatg tggtaattta cagtgcaatt cagttgctgg tccaaacact ttgtaacagt
     1681 ttaaagaatt gtggttacta gtaaatcctt aatttggcat catctaggtg tcattctttg
     1741 ccacttttga atcacacaaa tgaaccaaac aatggttagc tagcatcaaa catttaccaa
     1801 ataatctcat cagtgtcacc ccaatgttta agaagccaca ttttatgtcc ccagatatat
     1861 taatattatt ttatatataa tttaaaatag taatgtggtg ttactgtatt tgtcatataa
     1921 cgcaatatat acagtatgtc tttatctgta tcaattttgt ggtcacacca tcaataaact
     1981 ccagtttaat tatttaaaat tgggtagtga taaccatata aaggaacagt ggctagttcc
     2041 atgttgtagc tcccaacctg ctttcaggtg atccatgcag tgtcgtataa aaaggtaaca
     2101 atttttaacc ttggaaagca agtacactta aggtcccttt gctggtctcg acctagtgaa
     2161 gagcattttc tgtgtgtaga atagtgtgtt aggggtagct tctagactgg acctcggtgg
     2221 cttctaatcc gtgagtccta gcgggtgaca gccctccgct ttcacaggcg gaggagagtc
     2281 ctccgtaggg ttcctcggag tactgtcctc cggctagtgc gaccgtctat ataagcagag
     2341 ctggtttagt gaaccgtcag atccggaggc tgccaaccag ctccagcgga tccatgatgg
     2401 ggagtagcaa gagcaagcct aaagacccta gccagagaag caggggatct accggtgcat
     2461 ctgttgtagc tgatcttccg tcttatggac ctatcgcttt cgatcaggtg gatggcgtca
     2521 cgaacgcgac ccagctattc aaaaataata taaacaaaac cagaatgaac gatttaaacc
     2581 gcgaggagac acgattaaag accttcaccg actggccgct agattggctg gataaacgcc
     2641 agctggccca aaccggcatg tacttcacac acgctggcga caaagttaaa tgctttttct
     2701 gcggcgtgga aatcggttgc tgggagcagg aggatcagcc cgtgccggaa catcagcgat
     2761 ggtcgcccaa ctgtccactg ttgcgcagga gaactaccaa caatgtgccg atcaatgccg
     2821 aagcattaga tcgcatcctg ccgccaataa gctacgacat ctgcggcgcc aacgactcga
     2881 cgctagagat gaggagatct cccaagaaga ggaaggtgag atctgaagac ggcatggtga
     2941 gcaagggcga ggaggataac atggctatca tcaaggagtt catgcgcttc aaggttcaca
     3001 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3061 acgagggcac ccagaccgcc aagctgaagg tgacaaaggg cggccccctg cccttcgcct
     3121 gggacatcct gagccctcag ttcatgtacg gctccaaagc ctacgtgaag caccccgccg
     3181 acatccccga ctacttgaag ctgtctttcc cagagggctt caagtgggag cgcgtgatga
     3241 acttcgagga cggcggcgtg gtgacagtga cccaggactc ttctctgcag gacggcgagt
     3301 ttatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     3361 agaaaactat gggatgggag gcttcttccg agcggatgta cccagaggac ggagccctga
     3421 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgat gccgaggtca
     3481 agaccaccta caaggccaag aagcccgtgc aactgcccgg agcctacaac gtcaacatca
     3541 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3601 agggccgcca cagcaccggc ggcatggacg agctgtacaa gccgtacgga tctgccacca
     3661 attttagcct cctgaaacaa gcaggcgacg ttgaggagaa tccaggaccg tctcccatga
     3721 tggacgaggc ggaccgccag ctcctgagaa gatgcagggt gcgcctagtg agcgagctgc
     3781 aagtcgcgga gctgtgggac gctctgctga gtcgagagct gttcacgcgc gacatgatcg
     3841 aggatattca gcaggcagga tctggctctc ggcgggatca ggctaggcag ctcgtcacag
     3901 acctggagac tagaggccgc caggctctgc ctctgttcat ctcctgctta gaggacacag
     3961 gccaaggcac cctggcttca ctcttgcaaa gcggtcggca agcagccaag caagacccag
     4021 aggctgttaa acccctagac catctggtgc ctgtggtcct gggaccaatg ggactcacag
     4081 caaaggagca gagagtagtg aagctggacc cgtcacagcc tgccgtggga aacctgaccc
     4141 cagtggtgct cggacctgaa gaactctggc ctgctcgcct gaagccagag gttctcagac
     4201 cagaaacacc tagacccgtg gacattggtt ctggtggagc ccatgatgtc tgtgttccag
     4261 ggaagatcag gggacatgca gatatggcat acaccctgga ctctgatccc tgcggccact
     4321 gtctcatcat caacaatgtg aacttctgcc cttcctcggg gctcggcaca cgcacgggct
     4381 ccaacttgga ccgtgacaaa ctggagcacc gattccgctg gctgcgcttc atggtggagg
     4441 tgaagaacga cctgactgct aagaaaatgg tcacggcttt gatggagatg gcacaccgga
     4501 accaccgtgc cctggactgc tttgtggtgg tcatcctctc tcatggctgc caggccagcc
     4561 acctccagtt cccaggtgct gtttatggga cagatggatg ctccgtgtcc attgagaaaa
     4621 ttgtgaacat ctttaacgga agcggctgcc ctagcctggg agggaagccc aaactgttct
     4681 tcatccaggc ttgcggaggt gagcagaaag accacggctt tgaggtggct tgcacttcct
     4741 ctcagggcag aaccttggac agtgactctg agcctgatgc tgtcccctat caggaaggcc
     4801 caagaccctt ggaccaactg gatgctgtgt caagtttgcc tacccctagc gacatccttg
     4861 tgtcctactc caccttccca ggatttgtgt cttggaggga caagaaaagt ggctcctggt
     4921 acatcgagac tttggacggt attctggaac agtgggctcg ctctgaggat ctgcagtccc
     4981 tccttctcag ggttgccaat gctgtttctg cgaaagggac ttacaagcag attcctggct
     5041 gttttaactt cctgcggaaa aagctgtttt tcaaaacttc tgtgcccggg ccagagaacc
     5101 tgtacttcca ggggctcgag ccctctgccg gagacatgag agctgcaaac ctttggccga
     5161 gcccgctgat gattaaacgc tctaagaaga acagcctggc cttgtccctg accgccgacc
     5221 agatggtcag tgccttgttg gatgccgagc cccccatact ctattccgag tatgatccta
     5281 ccagaccctt cagtgaggct tcgatgatgg gcttactgac caacctggca gacagggagc
     5341 tggttcacat gattaactgg gccaagaggg tgccaggctt tgtggatttg accctccacg
     5401 atcaggtcca cctcctagaa tgtgcctggc tagagatcct gatgattggt ctcgtctggc
     5461 gcagcatgga gcacccagtg aagctactgt ttgctcctaa cttgctcttg gacaggaacc
     5521 agggaaaatg tgtagagggc atggtggaga ttttcgacat gctgctggct acatcatctc
     5581 ggttccgcat gatgaatctg cagggagagg aatttgtgtg cctcaaatct attattttgc
     5641 ttaattctgg agtttacaca tttctgtcca gcaccctgaa gtctctggag gaaaaggacc
     5701 atatccatcg agtgctggac aagatcacag acactttgat tcacctgatg gctaaggctg
     5761 gcctgaccct gcagcagcag catcagagac tcgcacagct tctcctgatc ctctcccaca
     5821 tcaggcacat gagtaacaaa ggcatggagc atctgtactc tatgaagtgc aagaacgtgg
     5881 tgcccctcta tgatctgctg ctggaggcgg cggacgctca ccgcctacat gcgcccacta
     5941 gccgtggagg ggcatccgtg gaggaaacag accaaagcca cttggctact gcgggctcta
     6001 cttcatctca ttccttgcaa aagtattaca tcacggggga ggcagagggt ttccctgcca
     6061 cagcaccagg atctacgtag acatgataag atacattgat gagtttggac aaaccacaac
     6121 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     6181 aaccatatta agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     6241 ggttcagggg gaggtgtggg aggttttttc ctgcaggact ctagtcgcgg ccgctaggga
     6301 taacagggta atggccgctc ttccgctaat actcaagtac aattttaatg gagtactttt
     6361 ttacttttac tcaagtaaga ttctagccag atacttttac ttttaattga gtaaaatttt
     6421 ccctaagtac ttgtactttc acttgagtaa aatttttgag tactttttac acctctgccc
     6481 tgcatcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8281 ccccgaaaag tgccacct
//
