LOCUS       sequence-227463-        6678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For broad or variegated gal4 inducible expression of Brainbow in 
            zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6678)
  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 6678)
  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-V
FEATURES             Location/Qualifiers
     source          1..6678
                     /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"
     primer_bind     complement(3216..3232)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3217..3233)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3257..3276)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3258..3276)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3309..3326)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3309..3325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3340)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3467..3922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3554..3573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3764..3785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3948..4052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4053..4121,4122..4913)
                     /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     complement(4271..4290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5084..5672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5573..5592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5826..5843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5960..5981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5996..6001,6002..6019,6020..6026)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6034..6050
                     /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)."
     primer_bind     6039..6061
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6058..6074
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6058..6074
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6118..6136
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     6173..6189
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6173..6189
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        6642..6660
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
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 gttaatgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     3241 cccggtaccc aattcgccct atagtgagtc gtattacgcg cgcattaccc tgttatccct
     3301 acgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3901 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4021 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4081 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4141 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4201 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4261 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4321 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4381 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4441 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4501 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4561 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4621 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4681 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4741 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4801 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4861 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4921 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4981 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5041 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5101 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5161 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5221 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5281 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5341 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5401 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5461 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5521 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5581 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5641 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5701 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5761 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5821 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5881 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5941 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6001 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6061 gaaacagcta tgaccatgat tacgccaagc gcgtagggat aacagggtaa tgcgcgcaat
     6121 taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc gctctagaac
     6181 tagtggatcc cccgggctgc aggaattcga tatcaagctt aggcctccaa ggcggagtac
     6241 tgtcctccgg gctggcggag tactgtcctc cggcaaggtc ggagtactgt cctccgacac
     6301 tagaggtcgg agtactgtcc tccgacgcaa ggcggagtac tgtcctccgg gctgcggagt
     6361 actgtcctcc ggcaaggtcg gagtactgtc ctccgacact agaggtcgga gtactgtcct
     6421 ccgacgcaag gtcggagtac tgtcctccga cactagaggt cggagtactg tcctccgacg
     6481 caaggcggag tactgtcctc cgggctggcg gagtactgtc ctccggcaag ggtcgactct
     6541 agagggtata taatggatcc catcgcgtct cagcctcact ttgagctcct ccacacgaat
     6601 tccctcgacc tcgaagacgc gtctgtttac cggtggcgcg caattaaccc tcactaaagg
     6661 gaacaaaagc tggagctc
//
