LOCUS       sequence-365950-        9146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible fish Hoxa13b activation via zebrafish transient 
            transgenesis or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9146)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 9146)
  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"
COMMENT     Sequence Label: pT2iU6m5Hoxa13bE4PCh(#765)
FEATURES             Location/Qualifiers
     source          1..9146
                     /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"
     misc_feature    join(179..190,191..378)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        3975..3993
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3975..3992
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4043..4072
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4082..4111
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4121..4150
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4160..4189
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4199..4228
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5144..5164
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6131..6187
                     /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             6197..6907
                     /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"
     polyA_signal    6935..7056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6946..6965)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7000..7019
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(7176..7313,7314..7325)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(7397..7985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7477..7496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8156..8947,8948..9016))
                     /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     8779..8798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9017..9121)
                     /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 tttttaggtc
      421 ccagatctga agctacccct aacacactat tctacacaca gaaaatgctc ttcactaggt
      481 cgagaccagc aaagttctag aatttgtcga aacatttatg ttatatattt cctgaaaaaa
      541 attctgagta agttcttaag tgttattgcc agcaacataa acaacagacg gcaaaatgaa
      601 taaatgataa caaagcagta ggcttaaata aacctaattt ttataggctg ttctctacaa
      661 ccctcaaaca gtgattagtt ttgtacttat aaacttgccc tttcattcat atttcaagaa
      721 aattggttca gaagatctgg atattctagc agttgttcaa gctcatggag ggatcagtga
      781 cctgattcca caatgactag gcctaatcca gaaattagat gactgtcaac ataaaaaggc
      841 acagcactca ctagctgccc tatatatttt attatatttt acatatatta ttttatttat
      901 ttagctctga gtgctgtact ttctggttaa agaaaactgc ttacaacagc taacctgtac
      961 tacctcaggc tcagggaatt tggaacaggt ttgtctggtt tgtttcttta accatgcatg
     1021 cttgttttca actatggcaa cacagtcaca tgggacatta cagaaatgat ttgtcgatga
     1081 catgcgactt ttctttaata aagcgcaaag atcccaaaaa gcaaactttt aacaaaaatc
     1141 atataattat attttcaatc cagctttgta gcaactttgt gctgctgttc actcagcaac
     1201 agatagtcag tataaggtca gtgtgtctca aagcagtgcc atctgtttca cacattgcgt
     1261 tctatatata agtgtgctgg ttgacacgac actgtataag gcctaggcta aaacacaaac
     1321 aatgtagaat gacactgtgt tttttttgta aacaaatgtt gtttttggtt aaacatcttt
     1381 gtgaaaacat cctcctgtca tgtatttgct atattcaaat gttaaacccg tgcagaatag
     1441 aacatataca aaaaaaaaca acacaacaca tttttaaaca ttattaaata tcaagtattg
     1501 ctggcagttc tgtttctgtt ttacagtacc ctttgccaca gttctccgct tttcctggtc
     1561 cagattccac aagtctgatt caccaatagc aaagcgaata aacaaccaaa gcagccaatc
     1621 actgcttgta gactgtcctg cgagaccggc ccattccagc acattctgga aacttccttt
     1681 atatgataat tataaataca tttaaattat tgatacaaaa catgtaattc ctagaacata
     1741 accatagcaa tcattagttt tcagggtaat tatgtatttt taggatttga ctgcggaaag
     1801 atctggtcat gtgacgtctc atgaacgtca cggccctggg tttctataaa tacagtagga
     1861 ctctcgacca tcggcagatt tttcgaagaa gaagatcagt ttcaggagcc gtactgttcc
     1921 gtttcaacgc aaatattaac ggtaagagcg aatttcctag tttgttttca tgccattctt
     1981 taaaaccata gcgtattact ttaattatag taaactttcg ctttctttat tacaagagac
     2041 gttttgtgtt gattctccgc ggacattttc ggtcagacaa tcagaaaatg accgcggagg
     2101 accagtaact tgcattacac gtaagttaaa tcttcgtgta ttaaaatggt taggttgtta
     2161 acgtcaaata ggttaccgtg tttgcgtgtg atcaggttgg ttttgttaga tttttgtcag
     2221 tatttttaat ttatttgttt tagtttattt attttttttg ctgaatcata gtttgtgaac
     2281 aaagaacccg gatgttacat acagtacagc cgccatgtta cagagagtta taacttaatc
     2341 attttaaaaa taattttgcc ttacttttag tttgtcatgt tgagaaatga ggaaatgtta
     2401 aaatgaggaa atatccaatt aatttaatat atcaaaataa tccatgatta caatgcactg
     2461 aactggagaa aattaagatg ttttctagtg tcatgaaaca aatgtaagag atgtacattg
     2521 tagatgtttt atgtcaagaa ttggctagtt gatgcagcat actggcgata ctcagttgta
     2581 ataacagtaa cgttacatgt taatagacta ctgagtatgc tgttctgtct atgtatgctc
     2641 tgtaagctac gagaaggact tttttaaaca gtaaagggtg caatattttt acaaattgaa
     2701 ttaaataaag gctgtctatt aagtaatatg cttgatattt ttcttacttg atcggaaata
     2761 agaaaaaata taaacgttgt tgctctaaaa atcctagttc agtttagcca accacaaata
     2821 cctttttgtt cctccaacag tttttttttc ttctctataa tatttggcag tctatagtac
     2881 tccaaatgtt tccccacagt ctaactaatt ggtacagcca aaatcatgac acttattgca
     2941 ataataattt tggttcattg gcattgttga tagcctgtgc cactaatatg gtcgattgat
     3001 catgcttcag gaagaaaact atattgtttg atgtaagatt attaaatctt cacctgcttc
     3061 cattacaaac tattcccatc ttattgaatt ctggtatgtc ttaaaggatt agttcacttc
     3121 ccaaatcaaa atttacttag ccttttttca tccatgatcc ctttttttca tcattaatga
     3181 agaaattgtt tttgaaaaag tttcaagatt tttttctcta tattgtggag cttgttagtt
     3241 taaaattcca aaatgcaata tgtggcttca aatggttcta aatgatccca gtcaaggaat
     3301 aacagtctta tctaatgaaa ccattagacc tttaaaaata aaaaaaataa aagtatttat
     3361 ttttaaatga ctgagtgatt aagttgaatt tcagcgtttc cttactgtgt agaagtcctt
     3421 ccttactggc cccacccttt ggttctctgc caatctgcta cctaatgtaa tgttgtggaa
     3481 cattattatt ctttatttct taatttttta ttttttattt taaaacaatg taaactgcac
     3541 agatgtgcag tttgtttaaa atggccaatg ctttggaaat gcatgacata attagatttc
     3601 atgatgcaca aagccaaatc tcagagcttg tgcaaaatga gctatcattt cactaggtaa
     3661 gaccctaaat tttcatatag gatcatttgg acaattttgc tgcaggtaaa atgcattcta
     3721 tagtccactg tcagccattg ttttggatag tatttatttt tctctacaag tatagtcaat
     3781 agttttctat tattttaaag gtttgtaaca tttaagggtg accaaatgca aagtaaaatt
     3841 tcattttcgg gtgaactatc tcgtttaaca tgggagaagt gcaaaacata cattattggc
     3901 tagaacattg tagtattttt taaatggaaa tgtgtgattg ctaatcttac tttgaatttg
     3961 tttacaggga tctgatttag gtgacactat agaatacaag ctacttgttc tttttgcagg
     4021 atcccatcga tttaaagcta tggagcaaaa gctcatttct gaagaggact tgaatgaaat
     4081 ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga
     4141 agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga
     4201 gcaaaagctc atttctgaag aggacttgaa tgaaatggag agcttgggcg acctcaccat
     4261 ggctagcaca gcgtctttac tcctccattc ccgctggatt gacccggtga tgtttctcta
     4321 tgacaacggt ttggatgata tgagcaaaaa catggaagga tttgtgggag gcaattttgc
     4381 tgcgaaccag tgtaggaatc tcattgccca tccgtctact ttggctccga gtacaaccta
     4441 cacatcgagt gaggtgccag tgtcgggcat gggagagcct gtcaaacaat gcagcccgtg
     4501 ttcggccgtg caaaataccc caagtgcttc tttgccctat ggatattttg gcggcagcta
     4561 ttacccgtgt aggatgccca agtcctgcac gcaacccacc acatatggag aaaaatacat
     4621 ggacacttct gtttctggag aggagtttcc ttctagggca aaggaatttg ctttctacca
     4681 gggctactcg tctggccctt accagcctgt accaagttac cttgacgtgc cagttgtgcc
     4741 tgctctgagc gcaccttcgg aaccgagaca tgaatctcta ctgcctgtgg aaacgtacca
     4801 gccgtgggca atcactaacg ggtggagcag tccagtgtac tgcccgaaag atcaaacaca
     4861 gtcaagtacc ttgtggaaat cgtctatcca agataccgtg tcaggaaccg atggagcctc
     4921 agtacgtcga gggagaaaga agcgggttcc ctacaccaag gttcaattga aggaactaga
     4981 acgagaatat gccacaaata agttcatcac aaaggacaaa cgaagaagga tctctgctca
     5041 cacaaacctg accgagcgac aagtgaccat ctggtttcag aacaggcggg taaaagaaaa
     5101 gaaagtggtc aacaagtaca agggcatcag ccaagctggg ccagagaacc tgtacttcca
     5161 ggggctcgag ccctctgccg gagacatgag agctgcaaac ctttggccga gcccgctgat
     5221 gattaaacgc tctaagaaga acagcctggc cttgtccctg accgccgacc agatggtcag
     5281 tgccttgttg gatgccgagc cccccatact ctattccgag tatgatccta ccagaccctt
     5341 cagtgaggct tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat
     5401 gattaactgg gccaagaggg tgccaggctt tgtggatttg accctccacg atcaggtcca
     5461 cctcctagaa tgtgcctggc tagagatcct gatgattggt ctcgtctggc gcagcatgga
     5521 gcacccagtg aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg
     5581 tgtagagggc atggtggaga ttttcgacat gctgctggct acatcatctc ggttccgcat
     5641 gatgaatctg cagggagagg aatttgtgtg cctcaaatct attattttgc ttaattctgg
     5701 agtttacaca tttctgtcca gcaccctgaa gtctctggag gaaaaggacc atatccatcg
     5761 agtgctggac aagatcacag acactttgat tcacctgatg gctaaggctg gcctgaccct
     5821 gcagcagcag catcagagac tcgcacagct tctcctgatc ctctcccaca tcaggcacat
     5881 gagtaacaaa ggcatggagc atctgtactc tatgaagtgc aagaacgtgg tgcccctcta
     5941 tgatctgctg ctggaggcgg cggacgctca ccgcctacat gcgcccacta gccgtggagg
     6001 ggcatccgtg gaggaaacag accaaagcca cttggctact gcgggctcta cttcatctca
     6061 ttccttgcaa aagtattaca tcacggggga ggcagagggt ttccctgcca cagcaccagg
     6121 ttccggcagc gctacaaact tctctctgct caagcaggct ggagatgtgg aagaaaaccc
     6181 tgggcccgaa gacggcatgg tgagcaaggg cgaggaggat aacatggcta tcatcaagga
     6241 gttcatgcgc ttcaaggttc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     6301 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgacaaa
     6361 gggcggcccc ctgcccttcg cctgggacat cctgagccct cagttcatgt acggctccaa
     6421 agcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtctt tcccagaggg
     6481 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgacag tgacccagga
     6541 ctcttctctg caggacggcg agtttatcta caaggtgaag ctgcgcggca ccaacttccc
     6601 ctccgacggc cccgtaatgc agaagaaaac tatgggatgg gaggcttctt ccgagcggat
     6661 gtacccagag gacggagccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     6721 cggccactac gatgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcaactgcc
     6781 cggagcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     6841 cgtggaacag tacgaacgcg ccgagggccg ccacagcacc ggcggcatgg acgagctgta
     6901 caagtaaggc gcgccgatat ctacgtagac atgataagat acattgatga gtttggacaa
     6961 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     7021 ttatttgtaa ccatattaag ctgcaataaa caagttaaca acaacaattg cattcatttt
     7081 atgtttcagg ttcaggggga ggtgtgggag gttttttcct gcaggactct agtcgcggcc
     7141 gctagggata acagggtaat ggccgctctt ccgctaatac tcaagtacaa ttttaatgga
     7201 gtactttttt acttttactc aagtaagatt ctagccagat acttttactt ttaattgagt
     7261 aaaattttcc ctaagtactt gtactttcac ttgagtaaaa tttttgagta ctttttacac
     7321 ctctgccctg catcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7801 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     8041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     8101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8281 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     9001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     9061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9121 gcacatttcc ccgaaaagtg ccacct
//
