LOCUS       sequence-227466-        7423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For broad gal4 inducible expression of Brainbow in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  AUTHORS   Pan YA, Freundlich T, Weissman TA, Schoppik D, Wang XC, Zimmerman S,
            Ciruna B, Sanes JR, Lichtman JW, Schier AF
  TITLE     Zebrabow: multispectral cell labeling for cell tracing and lineage 
            analysis in zebrafish.
  JOURNAL   Development. 2013 Jul;140(13):2835-46. doi: 10.1242/dev.094631.
  PUBMED    23757414
REFERENCE   2  (bases 1 to 7423)
  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"; doi: "10.1242/dev"; 
            journalName: "Development"; date: "2013-07"; volume: "140"; issue: 
            "13"; pages: "2835-46"
COMMENT     Sequence Label: UAS:Zebrabow-B
FEATURES             Location/Qualifiers
     source          1..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(43..76)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      89..98
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             95..796
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(228..246)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     496..515
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     690..713
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    926..1047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1130..1163)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1177..1186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1183..1185,1186..1188,1189..1902)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2025..2146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2153..2186)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2194..2203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2200..2202,2203..2205,2206..2919)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3536..3723,3724..3735)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3769..3785)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3769..3785)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3822..3842)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3822..3840)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3861..3877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3861..3877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3874..3896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3885..3901
                     /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(3909..3915,3916..3933,3934..3939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3954..3975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4092..4109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4263..4851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4343..4362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5022..5813,5814..5882))
                     /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     5645..5664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5883..5987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6150..6171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6521..6532,6533..6695)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    7129..7153
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7387..7411)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ataacttcgt ataggatact ttatacgaag ttataactcg agataacttc gtatagcata
       61 cattatacga agttataccg gcagatctgc caccatggtg agcaagggcg aggaggtcat
      121 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
      181 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      241 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
      301 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
      361 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
      421 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
      481 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
      541 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
      601 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
      661 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
      721 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat
      781 ggacgagctg tacaagtaaa gtgtgcggcc gcgactctag atcataatca gccataccac
      841 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      901 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      961 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1021 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat gctccgaggg
     1081 ccgccaccac ctgttcctgt aggaattggc cgcgactcta ggatctagga taacttcgta
     1141 taggatactt tatacgaagt tattgatcca ccggtcgcca ccatggtgag caagggcgag
     1201 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1261 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1321 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1381 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1441 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1501 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1561 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc
     1621 atcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1681 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1741 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1801 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1861 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     1921 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1981 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaatcg agataacttc
     2161 gtatagcata cattatacga agttataccg gtcgccacca tggtgagcaa gggcgaggag
     2221 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2281 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2341 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     2401 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2461 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2521 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2581 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2641 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2701 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2761 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2821 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2881 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     2941 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3001 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3181 gttaatcctt agggataaca gggtaatacg cgtgatctgc gaagatacgg ccacgggtgc
     3241 tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact
     3301 tcctccattt tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc
     3361 tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct
     3421 gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta
     3481 aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg
     3541 atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt
     3601 ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa
     3661 gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag
     3721 tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt
     3781 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     3841 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3901 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3961 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4021 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4081 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4141 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4201 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4561 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4681 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4921 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4981 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5041 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5101 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5161 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5221 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5281 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5341 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5401 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5461 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5521 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5581 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5641 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5701 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5761 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5821 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5881 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5941 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6001 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     6061 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6121 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6181 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccacgctggc
     6241 taagaactca tcagcctccc cggtccatct acccacgtac caatgcacca attggccaca
     6301 atgacggcta ctacatggtg ccattccttc ctctttatag gaatggagac tacctcctgt
     6361 ccaacaatgc tcttggatac gagtacgcct acctgttgga cccaggtcat tgcacaacac
     6421 cagaaatgcc ctctgatctg caaaagacgt gaataatctg ttcagacacc catatgcact
     6481 ctgttccaca caggtcagag gtttgtccag gagttcttga cagaggtgta aaaagtactc
     6541 aaaaatttta ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta
     6601 aaagtatctg gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta
     6661 cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa actagagatt cttgtttaag
     6721 cttttaatct caaaaaacat taaatgaaat gcatacaagg ttttatcctg ctttagaact
     6781 gtttgtattt aattatcaaa ctataagaca gacaatctaa tgccagtaca cgctactcaa
     6841 agttgtaaaa cctcagattt aacttcagta gaagctgatt ctcaaaattg ttagtgtcaa
     6901 gcctagctct tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc tcacaggcag
     6961 ccgatgcggg aagaggtgta ttagtcttga tagagaggct gcaaatagca ggaaacgtga
     7021 gcagagactc cctggtgtct gaaacacagg ccagatgggc cattcgtacg cttaatttcg
     7081 tatagggata acagggtaat gatatcgata tcaagggccc atctcgagac aagtttgtac
     7141 aaaaaagcag gctccgaatt cctgcagccc gggggatcac gcggccatca agcttaggcc
     7201 tccacggtgg cttctaatcc gtgagtccta gcgggtgaca gccctccgtc ttcacaggcg
     7261 gaggagagtc ttccgtaggg ttcctcggag tactgtcctc cgacgcgtgc aagggtcgac
     7321 tctagagggt atataatgga tcccatcgcg tctcagcctc actttgagct cctccacacg
     7381 aattcgaccc agctttcttg tacaaagtgg tgctagcgag ctc
//
