LOCUS       sequence-385019-        6735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P0.8 promoter for transgenic enhancer assays in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6735)
  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 6735)
  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-P0.8:EGFP;cryaa:mCherry-ISceI-Tol2
FEATURES             Location/Qualifiers
     source          1..6735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1519..1974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1656..1677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1868..1887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2101..2123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2115..2132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2116..2132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2142..2160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(2176..2187,2188..2350)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             join(3230..3232,3233..3235,3236..3949)
                     /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(3275..3296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3536..3555)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3883..3904
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3979..4113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4120..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4364..5074)
                     /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(4444..4467)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4642..4661)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4911..4929
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5071..5080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(5777..5964,5965..5976)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6010..6026)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6010..6026)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6063..6083)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6063..6081)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6102..6118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6102..6118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6115..6137)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6126..6142
                     /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(6150..6156,6157..6174,6175..6180))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6195..6216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6333..6350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6504..6735,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6584..6603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaaa tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1561 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1621 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     1681 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1741 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1801 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     1861 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     1921 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     1981 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2041 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2101 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     2161 gcgaattggg taccacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac
     2221 aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt
     2281 gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta
     2341 aaagcaagaa agaaaactag agattcttga agctttaggg ataacagggt aatctcgagc
     2401 tagcctaggc gccatcgatg tttaaacgga tcctgcttcg ggcttgttga aagggtcttc
     2461 tctagtcttc aagttcatga tggacacagg ggtgaaccag accagaccgg catcagggtg
     2521 attcactgga gaaatgagtc aatcatccac agctataaaa agggagattc ctgcttgagt
     2581 ttctcaatat gcacatcaac acaccagagg taagcgctca ctcattcttg tcaacatggg
     2641 caccaaacat ctctatatga cctttaggct tcttgactgt gtagtttcat ttattattgg
     2701 actgtactca gcaacttaag tgcattgatt tttggttttt tgaattacac acaactaagg
     2761 gactgattta gcatgaatct tcaatgtagt gaagatcata aatcattcta atcgtgctga
     2821 gttagtggtt aaataaacat atctaattgt tgaaagcaga cattttagtt gaaattatga
     2881 ttatttttaa ttattcttaa gttttaatgt tggttttgta ttgtctaatg attaaaatgt
     2941 ataaataaaa actaaaaact aaacataaaa aatagtatta ctttcatttt gtctaaggaa
     3001 tgtcaatttg tatttattta tttattttaa agtgcacaaa ataaaaatac atgtgtttga
     3061 gtaaaacctt atttaaagtg tatttaactg ttgtgtaaat gagtaatgtt taggctagct
     3121 ttattcatta ttttactgta tttaacatat tgaactggga tattctgatg tacacaacaa
     3181 actatttgaa aaaatataat ttcatttggt cttttgcaga aatgaattca tggtgagcaa
     3241 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3301 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3361 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3421 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3481 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3541 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3601 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3661 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3721 gaacttcaag atccgccaca acatcgagga cggcagcgcg cagctcgccg accactacca
     3781 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3841 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3901 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtagt ctagaactat
     3961 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     4021 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     4081 tttgtaacca ttataagctg caataaacaa gttcatatgt aagatacatt gatgagtttg
     4141 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     4201 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4261 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     4321 acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg tacagctcgt
     4381 ccatgccgcc ggtggagtgg tggccctcgg cgcgctcgta ctgttccacg atggtgtagt
     4441 cctcgttgtg ggaggtgatg tccagcttga tgttgacgtt gtaggcgccg ggcagctgca
     4501 cgggcttctt ggccttgtag gtggtcttga cctcggcgtc gtagtggccg ccgtccttca
     4561 gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac atccgctcgg
     4621 aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag gggaagttgg
     4681 tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag tcctgggtca
     4741 cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     4801 aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt
     4861 acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct
     4921 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     4981 actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga
     5041 tggccatgtt gtcatcctcg cccttgctca ccatggtggc gaccggtaat gtcagacctg
     5101 gtaactcctt actgtaactt attgcacctg atgaacaaaa gatgtgaact gaacgcaagt
     5161 aggaggctaa tagaggctag tacagtacaa agatccccaa atgatgtgtg ccggccttag
     5221 tggaggcaga cccctggtta tatacctgtg ctgcagaaag aggggcttac aaatggatct
     5281 ctctggaatg gcattatctc atggtgggaa cagtgttgtc agcagaatca gcggggtgga
     5341 ctctgacctg agaatgacag tctagcactg aatggctcag acaatactct tgtggccaac
     5401 tatcaccgga aaataagtgc ctgccaaaac aaaccagacc agatgaagct acatggctgt
     5461 agaacgtcag tatttaatgt gttgtggcca cttggttcac cagatttaag agctaatttt
     5521 tttaaatatc acagataatc aatcgctttt tttttttctt gactgaaaag cttagactgg
     5581 aaaattcttt gaaaaagttt gcactacaga aaataaatac ataacatcct gcactgaatg
     5641 cacactatta atagaaaaga gagcattatt gtaggaaatg tattgtatat gtacattctg
     5701 tgcatttcac aattttgggc ccgtagggat aacagggtaa tcaattggtt tggtaatagc
     5761 aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat
     5821 tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta
     5881 ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt
     5941 aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct accttcagca
     6001 agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt
     6061 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6121 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6181 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6241 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6301 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6361 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6421 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6721 agttcggtgt aggtc
//
