LOCUS       sequence-178818-        7899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains She enhancer sequence, and She specific gene promoter 
            driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
  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 7899)
  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-P1Egfp
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     630..646
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    666..686
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1133..1157
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1908..1917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1914..1916,1917..1919,1920..2630)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1914..1916,1917..1919,1920..2630)
                     /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(1959..1980)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2220..2239)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2567..2588
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2631..2696
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2717..2741)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2767..2787)
                     /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(2795..2811)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2827..2961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3400..3587,3588..3599)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3633..3649)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3649)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3686..3706)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3686..3704)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3725..3741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3738..3760)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3749..3765
                     /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(3773..3779,3780..3797,3798..3803))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3818..3839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3956..3973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4886..5677,5678..5746))
                     /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     5509..5528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5747..5851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6014..6035)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6384..6395,6396..6558)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6927..7048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7171..7881)
                     /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(7251..7274)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7449..7468)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7718..7736
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7878..7887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cagacctggt aactccttac tgtaacttat tgcacctgat gaacaaaaga tgtgaactga
       61 acgcaagtag gaggctaata gaggctagta cagtacaaag atccccaaat gatgtgtgcc
      121 ggccttagtg gaggcagacc cctggttata tacctgtgct gcagaaagag gggcttacaa
      181 atggatctct ctggaatggc attatctcat ggtgggaaca gtgttgtcag cagaatcagc
      241 ggggtggact ctgacctgag aatgacagtc tagcactgaa tggctcagac aatactcttg
      301 tggccaacta tcaccggaaa ataagtgcct gccaaaacaa accagaccag atgaagctac
      361 atggctgtag aacgtcagta tttaatgtgt tgtggccact tggttcacca gatttaagag
      421 ctaatttttt taaatatcac agataatcaa tcgctttttt ttttcttgac tgaaaagctt
      481 agactggaaa attctttgaa aaagtttgca ctacagaaaa taaatacata acatcctgca
      541 ctgaatgcac actattaata gaaaagagag cattattgta ggaaatgtat tgtatatgta
      601 cattctgtgc atttcacaat tttctcgagc aggaaacagc tatgaccatg attacgccaa
      661 gctatcaact ttgtatagaa aagttgtttg tttaagcaat atcttccttt gctccctctt
      721 ctctctgtcg tatctctctt tctctccaca tattctcttc tcattcaaca gctgtattat
      781 aagcagacac agtggagctg aaaagcacca aacacaatgt gagccctttc cttgcctctt
      841 cctgtctctt ttcctagtca tgaacacact cttcactctg tctttgtggg aaatacactt
      901 ttgtgctaca cacacacaca cacacccttg ctcacaaata cgccaagaat acaatgcata
      961 cataacacaa tggctctgtc ccattcttcc agtctctggt cagagcagtg tttgaagaat
     1021 tatgtcacct gatacatagg ctgtatcctc tcccatttac agacatccct tccagctaca
     1081 ctttgaacat aatagcacag aaatcagaat ctcatgaggt cacacacaca catcaagttt
     1141 gtacaaaaaa gcaggctcgc taaccacttg aaaattctaa tcttttttcc atattttaat
     1201 taaaaagctg tcatatgttt ttattatatc actgaaaatc agggttgttt ctaaacacct
     1261 attcttaact gttctgaagt ttaaactgca tggtctaaaa atgaacagaa acatcgccct
     1321 ttttttctgt cttatatggt tgtacgtgtg tacctgcgtg cgcgtgtgtt ctttaatatg
     1381 taattttttg tttatgtgtg tttatttttc aaagtcttgc acaaacttct tagcactctt
     1441 ttcccctgaa actactagct gatgtttttt aatcacataa gaatttattt agcccacata
     1501 taattgtttt gctctttatt gttagcactt aaagtgcttt ataaattaaa taaaattgaa
     1561 tcattttttg cgctgattta acttaatcat aatcaattaa actaaagggt tcaaatgcgc
     1621 ctcaacaggt gtccagcgaa taagaaagtt gccattgagg ggaacagaag ctgcagaggt
     1681 gcagaaacca cccccacccg cagctgtagc tgcagcacaa aagcacgtac atccacatcc
     1741 ccttcacttc atctagactg tcgccaggag tggatatacc gggaatggga tcttcagcga
     1801 gagaaaagaa aaacttcaaa ttcttccgcg tttatgccct tcagaggctc ctctttatct
     1861 tgcgggaata attgggagca aactaaaaca agacaataca ttccatcgcc accatggtga
     1921 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1981 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2041 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2101 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2161 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2221 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2281 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2341 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     2401 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2461 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2521 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2581 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
     2641 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     2701 gatctagata actgataccc agctttcttg tacaaagtgg ggcgcgccta cgtaactagt
     2761 ctcgagcaac tttattatac atagttgata attcactggc cgtcgtttta cggtaccatc
     2821 gatgatgatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2881 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2941 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3001 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     3061 tatgatcctc tagatcagat ctgcgaagat acggccacgg gtgctcttga tcctgtggct
     3121 gattttggac tgtgctgctc gcagctgctg atgaatcaca tacttcctcc attttcttcc
     3181 actgattgac tgttataatt tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat
     3241 agatgtgaca tgacgtcact tccaaaggac caatgaacat gtctgaccaa tttcatataa
     3301 tgtgaaaacg attttcatag gcagaataaa taacatttaa attaaactgg gcatcagcgc
     3361 aattcaattg gtttggtaat agcaagggaa aatagaatga agtgatctcc aaaaaataag
     3421 tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat
     3481 gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc
     3541 caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt acacctctgg
     3601 ttcttgaccc cctaccttca gcaagcccag cagatccact agttctagag cggccgccac
     3661 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     3721 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3781 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3841 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3901 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3961 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4021 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4741 ggtctgacgc tcagtggaac gaaaactcac gttaaggcat tttggtcatg agattatcaa
     4801 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4861 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4921 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4981 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5041 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5101 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5161 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5221 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5281 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5341 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5401 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5461 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5521 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5581 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5641 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5701 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5761 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5821 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     5881 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5941 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     6001 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     6061 caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa ctcatcagcc
     6121 tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg gctactacat
     6181 ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca atgctcttgg
     6241 atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa tgccctctga
     6301 tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc acacaggtca
     6361 gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt
     6421 gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa
     6481 tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag
     6541 taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa
     6601 cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc
     6661 aaactataag acagacaatc taatgccagt acacgctact caaagttgta aaacctcaga
     6721 tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg
     6781 ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt
     6841 gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg
     6901 tctgaaacac aggcccattt acgccttaag atacattgat gagtttggac aaaccacaac
     6961 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     7021 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     7081 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     7141 ggctgattat gatctagagt cgcggccgct ttacttgtac agctcgtcca tgccgccggt
     7201 ggagtggtgg ccctcggcgc gctcgtactg ttccacgatg gtgtagtcct cgttgtggga
     7261 ggtgatgtcc agcttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc
     7321 cttgtaggtg gtcttgacct cggcgtcgta gtggccgccg tccttcagct tcagcctctg
     7381 cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca
     7441 gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt
     7501 caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc
     7561 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa
     7621 gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg
     7681 ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt
     7741 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     7801 cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg ccatgttgtc
     7861 atcctcgccc ttgctcacca tggtggcgac cggtaatgt
//
