LOCUS       sequence-178709-        7456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains She enhancer sequence and a basal promoter driving 
            expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  AUTHORS   Quillien A, Abdalla M, Yu J, Ou J, Zhu LJ, Lawson ND
  TITLE     Robust Identification of Developmentally Active Endothelial 
            Enhancers in Zebrafish Using FANS-Assisted ATAC-Seq.
  JOURNAL   Cell Rep. 2017 Jul 18;20(3):709-720. doi: 
            10.1016/j.celrep.2017.06.070.
  PUBMED    28723572
REFERENCE   2  (bases 1 to 7456)
  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: 
            "10.1016/j.celrep.2017.06"; journalName: "Cell Rep"; date: 
            "2017-07-18- 18"; volume: "20"; issue: "3"; pages: "709-720"
COMMENT     Sequence Label: pT2-cryR;sheCE1-basEgfp
FEATURES             Location/Qualifiers
     source          1..7456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     640..656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    676..696
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1143..1167
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1244..1253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1250..1252,1253..1255,1256..1966)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1250..1252,1253..1255,1256..1966)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1295..1316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1556..1575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1903..1924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1967..2032
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2156..2277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2284..2308)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2334..2354)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2362..2378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2394..2528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2967..3154,3155..3166)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3200..3216)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3200..3216)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3253..3273)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3253..3271)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3292..3308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3305..3327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3316..3332
                     /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(3340..3346,3347..3364,3365..3370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3385..3406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3523..3540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3694..4282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3774..3793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4453..5244,5245..5313))
                     /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     5076..5095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5314..5418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5581..5602)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5951..5962,5963..6125)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6494..6615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6738..7448)
                     /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(6818..6841)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7016..7035)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7285..7303
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7445..7454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ccggtaatgt cagacctggt aactccttac tgtaacttat tgcacctgat gaacaaaaga
       61 tgtgaactga acgcaagtag gaggctaata gaggctagta cagtacaaag atccccaaat
      121 gatgtgtgcc ggccttagtg gaggcagacc cctggttata tacctgtgct gcagaaagag
      181 gggcttacaa atggatctct ctggaatggc attatctcat ggtgggaaca gtgttgtcag
      241 cagaatcagc ggggtggact ctgacctgag aatgacagtc tagcactgaa tggctcagac
      301 aatactcttg tggccaacta tcaccggaaa ataagtgcct gccaaaacaa accagaccag
      361 atgaagctac atggctgtag aacgtcagta tttaatgtgt tgtggccact tggttcacca
      421 gatttaagag ctaatttttt taaatatcac agataatcaa tcgctttttt ttttcttgac
      481 tgaaaagctt agactggaaa attctttgaa aaagtttgca ctacagaaaa taaatacata
      541 acatcctgca ctgaatgcac actattaata gaaaagagag cattattgta ggaaatgtat
      601 tgtatatgta cattctgtgc atttcacaat tttctcgagc aggaaacagc tatgaccatg
      661 attacgccaa gctatcaact ttgtatagaa aagttgtttg tttaagcaat atcttccttt
      721 gctccctctt ctctctgtcg tatctctctt tctctccaca tattctcttc tcattcaaca
      781 gctgtattat aagcagacac agtggagctg aaaagcacca aacacaatgt gagccctttc
      841 cttgcctctt cctgtctctt ttcctagtca tgaacacact cttcactctg tctttgtggg
      901 aaatacactt ttgtgctaca cacacacaca cacacccttg ctcacaaata cgccaagaat
      961 acaatgcata cataacacaa tggctctgtc ccattcttcc agtctctggt cagagcagtg
     1021 tttgaagaat tatgtcacct gatacatagg ctgtatcctc tcccatttac agacatccct
     1081 tccagctaca ctttgaacat aatagcacag aaatcagaat ctcatgaggt cacacacaca
     1141 catcaagttt gtacaaaaaa gcaggctaga ctctagaggg tatataatgg atctcgcgtc
     1201 tcagcctcac tttgagctcc tccacacgct agcgctaccg gtcgccacca tggtgagcaa
     1261 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1321 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1381 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1441 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1501 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1561 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1621 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1681 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1741 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1801 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1861 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1921 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     1981 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     2041 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2101 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     2281 cgtacccagc tttcttgtac aaagtggggc gcgcctacgt aactagtctc gagcaacttt
     2341 attatacata gttgataatt cactggccgt cgttttacgg taccatcgat gatgatccag
     2401 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2461 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2521 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2581 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag
     2641 atcagatctg cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt
     2701 gctgctcgca gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt
     2761 tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga
     2821 cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt
     2881 ttcataggca gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt
     2941 tggtaatagc aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg
     3001 taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa
     3061 agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact
     3121 taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct
     3181 accttcagca agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc
     3241 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3301 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3361 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3421 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3481 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3541 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3601 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4321 gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5461 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5521 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5581 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5641 accgtctatc agggcgatgg cccacgctgg ctaagaactc atcagcctcc ccggtccatc
     5701 tacccacgta ccaatgcacc aattggccac aatgacggct actacatggt gccattcctt
     5761 cctctttata ggaatggaga ctacctcctg tccaacaatg ctcttggata cgagtacgcc
     5821 tacctgttgg acccaggtca ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg
     5881 tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag gtttgtccag
     5941 gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta
     6001 cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct tacttgagta
     6061 aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc
     6121 aagaaagaaa actagagatt cttgtttaag cttttaatct caaaaaacat taaatgaaat
     6181 gcatacaagg ttttatcctg ctttagaact gtttgtattt aattatcaaa ctataagaca
     6241 gacaatctaa tgccagtaca cgctactcaa agttgtaaaa cctcagattt aacttcagta
     6301 gaagctgatt ctcaaaattg ttagtgtcaa gcctagctct tttggggctg aaaagcaatc
     6361 ctgcagtgct gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta ttagtcttga
     6421 tagagaggct gcaaatagca ggaaacgtga gcagagactc cctggtgtct gaaacacagg
     6481 cccatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     6541 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     6601 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     6661 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     6721 ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgccggtgga gtggtggccc
     6781 tcggcgcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc
     6841 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc
     6901 ttgacctcgg cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc
     6961 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc
     7021 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg
     7081 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag
     7141 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg
     7201 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc
     7261 caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg
     7321 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc
     7381 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttgtcatc ctcgcccttg
     7441 ctcaccatgg tggcga
//
