LOCUS       sequence-383073-        7208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZSP promoter for transgenic enhancer assays in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  AUTHORS   Begeman IJ, Emery B, Kurth A, Kang J
  TITLE     Regeneration and developmental enhancers are differentially 
            compatible with minimal promoters.
  JOURNAL   Dev Biol. 2022 Sep 24;492:47-58. doi: 10.1016/j.ydbio.2022.09.007.
  PUBMED    36167150
REFERENCE   2  (bases 1 to 7208)
  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: "Dev Biol.";
            date: "2022-09-24"; volume: "492:47-58. doi"; pages: " 
            10.1016/j.ydbio.2022.09.007"
COMMENT     Sequence Label: Tol2-ISceI-ZSP:EGFP;cryaa:mCherry-ISceI-Tol2
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     66..83
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(127..138,139..301)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     intron          976..1548
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1622..1631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1628..1630,1631..1633,1634..2347)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1628..1630,1631..1633,1634..2344)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1673..1694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1934..1953)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2281..2302
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2403..2537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2544..2665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2788..3498)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGHHSTGGMDELYK"
     primer_bind     complement(2868..2891)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3066..3085)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3335..3353
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3495..3504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(4201..4388,4389..4400)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4434..4450)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4450)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4487..4507)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4487..4505)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4526..4542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4526..4542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4539..4561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4550..4566
                     /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(4574..4580,4581..4598,4599..4604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4619..4640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4757..4774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4928..5516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5008..5027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5687..6478,6479..6547))
                     /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     6310..6329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6548..6652)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6678..7133)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6815..6836)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7027..7046
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      121 gtaccacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta caagtactta
      181 gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact tgagtaaaag
      241 taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt aaaagcaaga
      301 aagaaaacta gagattcttg aagctttagg gataacaggg taatctcgag ctagcctagg
      361 cgccatcgat gtttaaacgg atcctgcaga tctttcgggc ttgttgaaag ggtcttctct
      421 agtcttcaag ttcatgatgg acacaggggt gaaccagacc agaccggcat cagggtgatt
      481 cactggagaa atgagtcaat catccacagc tataaaaagg gagattcctg cttgagtttc
      541 tcaatatgca catcaacaca ccagaggtaa gcgctcactc attcttgtca acatgggcac
      601 caaacatctc tatatgacct ttaggcttct tgactgtgta gtttcattta ttattggact
      661 gtactcagca acttaagtgc attgattttt ggttttttga attacacaca actaagggac
      721 tgatttagca tgaatcttca atgtagtgaa gatcataaat cattctaatc gtgctgagtt
      781 agtggttaaa taaacatatc taattgttga aagcagacat tttagttgaa attatgatta
      841 tttttaatta ttcttaagtt ttaatgttgg ttttgtattg tctaatgatt aaaatgtata
      901 aataaaaact aaaaactaaa cataatcact ttgagctcct ccacacgaat ttcgaccgat
      961 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
     1021 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
     1081 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
     1141 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
     1201 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
     1261 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
     1321 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
     1381 tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat cctgcctttc
     1441 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
     1501 ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct cctgggcaac
     1561 gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcctcgacgg atccaccggt
     1621 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1681 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1741 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1801 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1861 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1921 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1981 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2041 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     2101 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     2161 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2221 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2281 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2341 caagtaagga agttaaagca tgcgaattaa ttcatcgacc cgggaagagt tagatcgatg
     2401 atgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2461 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2521 gctgcaataa acaagttcat atgtaagata cattgatgag tttggacaaa ccacaactag
     2581 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2641 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2701 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     2761 tgattatgat ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgccggtgga
     2821 gtggtggccc tcggcgcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     2881 gatgtccagc ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     2941 gtaggtggtc ttgacctcgg cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     3001 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     3061 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     3121 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     3181 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     3241 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     3301 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg
     3361 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     3421 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttgtcatc
     3481 ctcgcccttg ctcaccatgg tggcgaccgg taatgtcaga cctggtaact ccttactgta
     3541 acttattgca cctgatgaac aaaagatgtg aactgaacgc aagtaggagg ctaatagagg
     3601 ctagtacagt acaaagatcc ccaaatgatg tgtgccggcc ttagtggagg cagacccctg
     3661 gttatatacc tgtgctgcag aaagaggggc ttacaaatgg atctctctgg aatggcatta
     3721 tctcatggtg ggaacagtgt tgtcagcaga atcagcgggg tggactctga cctgagaatg
     3781 acagtctagc actgaatggc tcagacaata ctcttgtggc caactatcac cggaaaataa
     3841 gtgcctgcca aaacaaacca gaccagatga agctacatgg ctgtagaacg tcagtattta
     3901 atgtgttgtg gccacttggt tcaccagatt taagagctaa tttttttaaa tatcacagat
     3961 aatcaatcgc tttttttttt tcttgactga aaagcttaga ctggaaaatt ctttgaaaaa
     4021 gtttgcacta cagaaaataa atacataaca tcctgcactg aatgcacact attaatagaa
     4081 aagagagcat tattgtagga aatgtattgt atatgtacat tctgtgcatt tcacaatttt
     4141 gggcccgtag ggataacagg gtaatcaatt ggtttggtaa tagcaaggga aaatagaatg
     4201 aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag
     4261 tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta
     4321 ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac
     4381 tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca gcagatccac
     4441 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     4501 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4561 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4621 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4741 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4801 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5101 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5341 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5641 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5701 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5761 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5821 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5881 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5941 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6001 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6061 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6121 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6181 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6241 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6301 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6361 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6421 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6481 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6541 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6661 cccgaaaaat gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     6721 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     6781 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     6841 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     6901 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     6961 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7021 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     7081 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     7141 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7201 ccagctgg
//
