LOCUS       sequence-178963-        7619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains She specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7619)
  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 7619)
  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;sheP1Egfp
FEATURES             Location/Qualifiers
     source          1..7619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..31
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      782..791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(788..790,791..793,794..1504)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(788..790,791..793,794..1504)
                     /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(833..854)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1094..1113)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1441..1462
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1505..1570
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1591..1615)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1641..1661)
                     /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(1669..1685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1701..1835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2274..2461,2462..2473)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2507..2523)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2560..2578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2599..2615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2612..2634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2623..2639
                     /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(2647..2653,2654..2671,2672..2677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2692..2713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2830..2847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3001..3589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3081..3100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3760..4551,4552..4620))
                     /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     4383..4402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4621..4725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4888..4909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5258..5269,5270..5432)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5801..5922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6045..6755)
                     /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(6125..6148)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6323..6342)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6592..6610
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6752..6761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     7403..7419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7439..7459
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7461..7480
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7461..7479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7488..7595
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7504..7520
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7505..7521
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7555..7571)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7608..7619,1..7))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gttaattcaa gtttgtacaa aaaagcaggc tcgctaacca cttgaaaatt ctaatctttt
       61 ttccatattt taattaaaaa gctgtcatat gtttttatta tatcactgaa aatcagggtt
      121 gtttctaaac acctattctt aactgttctg aagtttaaac tgcatggtct aaaaatgaac
      181 agaaacatcg cccttttttt ctgtcttata tggttgtacg tgtgtacctg cgtgcgcgtg
      241 tgttctttaa tatgtaattt tttgtttatg tgtgtttatt tttcaaagtc ttgcacaaac
      301 ttcttagcac tcttttcccc tgaaactact agctgatgtt ttttaatcac ataagaattt
      361 atttagccca catataattg ttttgctctt tattgttagc acttaaagtg ctttataaat
      421 taaataaaat tgaatcattt tttgcgctga tttaacttaa tcataatcaa ttaaactaaa
      481 gggttcaaat gcgcctcaac aggtgtccag cgaataagaa agttgccatt gaggggaaca
      541 gaagctgcag aggtgcagaa accaccccca cccgcagctg tagctgcagc acaaaagcac
      601 gtacatccac atccccttca cttcatctag actgtcgcca ggagtggata taccgggaat
      661 gggatcttca gcgagagaaa agaaaaactt caaattcttc cgcgtttatg cccttcagag
      721 gctcctcttt atcttgcggg aataattggg agcaaactaa aacaagacaa tacattccat
      781 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      841 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      901 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      961 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1021 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1081 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1141 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1201 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1261 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1321 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1381 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1441 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1501 caagtccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg
     1561 cccgggatcc accggatcta gataactgat acccagcttt cttgtacaaa gtggggcgcg
     1621 cctacgtaac tagtctcgag caactttatt atacatagtt gataattcac tggccgtcgt
     1681 tttacggtac catcgatgat gatccagaca tgataagata cattgatgag tttggacaaa
     1741 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1801 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1861 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1921 gtggtatggc tgattatgat cctctagatc agatctgcga agatacggcc acgggtgctc
     1981 ttgatcctgt ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc
     2041 ctccattttc ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg
     2101 tgcattgtgg taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga
     2161 ccaatttcat ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa
     2221 ctgggcatca gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat
     2281 ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt
     2341 ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt
     2401 aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta
     2461 ctttacacct ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct
     2521 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     2581 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag gcattttggt
     3661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4741 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4801 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4861 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4921 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc acgctggcta
     4981 agaactcatc agcctccccg gtccatctac ccacgtacca atgcaccaat tggccacaat
     5041 gacggctact acatggtgcc attccttcct ctttatagga atggagacta cctcctgtcc
     5101 aacaatgctc ttggatacga gtacgcctac ctgttggacc caggtcattg cacaacacca
     5161 gaaatgccct ctgatctgca aaagacgtga atatctgttc agacacccat atccactctg
     5221 ttccacacag gtcagaggtt tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa
     5281 aattttactc aagtgaaagt acaagtactt agggaaaatt ttactcaatt aaaagtaaaa
     5341 gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt actccattaa aattgtactt
     5401 gagtattaag gaagtaaaag taaaagcaag aaagaaaact agagattctt gtttaagctt
     5461 ttaatctcaa aaaacattaa atgaaatgca tacaaggttt tatcctgctt tagaactgtt
     5521 tgtatttaat tatcaaacta taagacagac aatctaatgc cagtacacgc tactcaaagt
     5581 tgtaaaacct cagatttaac ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc
     5641 tagctctttt ggggctgaaa agcaatcctg cagtgctgaa aagcctctca caggcagccg
     5701 atgcgggaag aggtgtatta gtcttgatag agaggctgca aatagcagga aacgtgagca
     5761 gagactccct ggtgtctgaa acacaggccc atttacgcct taagatacat tgatgagttt
     5821 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     5881 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     5941 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     6001 tacaaatgtg gtatggctga ttatgatcta gagtcgcggc cgctttactt gtacagctcg
     6061 tccatgccgc cggtggagtg gtggccctcg gcgcgctcgt actgttccac gatggtgtag
     6121 tcctcgttgt gggaggtgat gtccagcttg atgttgacgt tgtaggcgcc gggcagctgc
     6181 acgggcttct tggccttgta ggtggtcttg acctcggcgt cgtagtggcc gccgtccttc
     6241 agcttcagcc tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg
     6301 gaggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg
     6361 gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc
     6421 acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     6481 aaggacagct tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg
     6541 tacatgaact gaggggacag gatgtcccag gcgaagggca gggggccgcc cttggtcacc
     6601 ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     6661 aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg
     6721 atggccatgt tgtcatcctc gcccttgctc accatggtgg cgaccggtaa tgtcagacct
     6781 ggtaactcct tactgtaact tattgcacct gatgaacaaa agatgtgaac tgaacgcaag
     6841 taggaggcta atagaggcta gtacagtaca aagatcccca aatgatgtgt gccggcctta
     6901 gtggaggcag acccctggtt atatacctgt gctgcagaaa gaggggctta caaatggatc
     6961 tctctggaat ggcattatct catggtggga acagtgttgt cagcagaatc agcggggtgg
     7021 actctgacct gagaatgaca gtctagcact gaatggctca gacaatactc ttgtggccaa
     7081 ctatcaccgg aaaataagtg cctgccaaaa caaaccagac cagatgaagc tacatggctg
     7141 tagaacgtca gtatttaatg tgttgtggcc acttggttca ccagatttaa gagctaattt
     7201 ttttaaatat cacagataat caatcgcttt tttttttctt gactgaaaag cttagactgg
     7261 aaaattcttt gaaaaagttt gcactacaga aaataaatac ataacatcct gcactgaatg
     7321 cacactatta atagaaaaga gagcattatt gtaggaaatg tattgtatat gtacattctg
     7381 tgcatttcac aattttctcg agcaggaaac agctatgacc atgattacgc caagctatca
     7441 actttgtata gaaaagttgg taatacgact cactataggg cgaattgggt accgggcccc
     7501 ccctcgaggt cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatcca
     7561 ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgagg
//
