LOCUS       sequence-385059-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DSCP promoter for transgenic enhancer assays in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  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 6099)
  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-DSCP:EGFP;cryaa:mCherry-ISceI-Tol2
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     359..376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    493..514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(529..534,535..552,553..559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    567..583
                     /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     572..594
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     591..607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     591..607
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     626..646
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        628..646
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     683..699
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     683..699
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(733..744,745..932)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     regulatory      1629..1638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1635..2345
                     /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(1780..1798)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2048..2067
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2242..2265
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2468..2589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2596..2730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(2760..3473,3474..3476,3477..3479))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSAQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2805..2826)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3154..3173
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3413..3434
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(3723..3885,3886..3897)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        complement(3913..3931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3941..3958)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3941..3957)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3950..3972)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4099..4554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4186..4205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4396..4417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4580..4684
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4685..4753,4754..5545)
                     /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(4903..4922)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5716..6099,1..205)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
       61 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      121 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      181 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      241 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      301 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      361 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      421 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      481 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      541 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      601 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
      661 gctccaccgc ggtggcggcc gctctagaac tagtggatct gctgggcttg ctgaaggtag
      721 ggggtcaaga accagaggtg taaagtactt gagtaatttt acttgattac tgtacttaag
      781 tattattttt ggggattttt actttacttg agtacaatta aaaatcaata cttttacttt
      841 tacttaatta cattttttta gaaaaaaaag tactttttac tccttacaat tttatttaca
      901 gtcaaaaagt acttattttt tggagatcac ttcattctat tttcccttgc tattaccaaa
      961 ccaattgatt accctgttat ccctacgggc ccaaaattgt gaaatgcaca gaatgtacat
     1021 atacaataca tttcctacaa taatgctctc ttttctatta atagtgtgca ttcagtgcag
     1081 gatgttatgt atttattttc tgtagtgcaa actttttcaa agaattttcc agtctaagct
     1141 tttcagtcaa gaaaaaaaaa aagcgattga ttatctgtga tatttaaaaa aattagctct
     1201 taaatctggt gaaccaagtg gccacaacac attaaatact gacgttctac agccatgtag
     1261 cttcatctgg tctggtttgt tttggcaggc acttattttc cggtgatagt tggccacaag
     1321 agtattgtct gagccattca gtgctagact gtcattctca ggtcagagtc caccccgctg
     1381 attctgctga caacactgtt cccaccatga gataatgcca ttccagagag atccatttgt
     1441 aagcccctct ttctgcagca caggtatata accaggggtc tgcctccact aaggccggca
     1501 cacatcattt ggggatcttt gtactgtact agcctctatt agcctcctac ttgcgttcag
     1561 ttcacatctt ttgttcatca ggtgcaataa gttacagtaa ggagttacca ggtctgacat
     1621 taccggtcgc caccatggtg agcaagggcg aggatgacaa catggccatc atcaaggagt
     1681 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1741 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1801 gcggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1861 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1921 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1981 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     2041 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     2101 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2161 gccactacga cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2221 gcgcctacaa cgtcaacatc aagctggaca tcacctccca caacgaggac tacaccatcg
     2281 tggaacagta cgagcgcgcc gagggccacc actccaccgg cggcatggac gagctgtaca
     2341 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2401 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2461 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2581 tgtatcttac atatgaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2641 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2701 ctcatcaatg tatcttatca tgtctggatc tacgtaatac gactcactat agttctagac
     2761 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2821 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     2881 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2941 gcgagctgcg cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     3001 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     3061 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     3121 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     3181 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     3241 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     3301 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     3361 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3421 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     3481 ccgtttggta tgcgtcttgt gattcaaagt tggcttattc aaaggatatt aacgaaggca
     3541 gcggcacgtc tgctcggctc gaacattcat tcacaactga tgctctcagc caccccgcgc
     3601 ccttttatac cgctgcgctc gatccccggg cgagctcgag gatccgttta aacatcgatg
     3661 gcgcctaggc tagctcgaga ttaccctgtt atccctaaag cttcaagaat ctctagtttt
     3721 ctttcttgct tttactttta cttccttaat actcaagtac aattttaatg gagtactttt
     3781 ttacttttac tcaagtaaga ttctagccag atacttttac ttttaattga gtaaaatttt
     3841 ccctaagtac ttgtactttc acttgagtaa aatttttgag tactttttac acctctgtgg
     3901 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     3961 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4021 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4081 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4441 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4561 catttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4621 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4681 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4741 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4801 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4861 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4921 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4981 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5041 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5101 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5161 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5221 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5281 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5341 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5401 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5461 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5521 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5581 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5641 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5701 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5761 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5821 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5881 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5941 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6001 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6061 cttggagcga acgacctaca ccgaactgag atacctaca
//
