LOCUS       sequence-178971-        7567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains yrk specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  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 7567)
  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;yrkP1Egfp
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..26
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      725..734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(731..733,734..736,737..1447)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(731..733,734..736,737..1447)
                     /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(776..797)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1037..1056)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1384..1405
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1448..1513
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1534..1558)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1584..1604)
                     /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(1612..1628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1644..1778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2217..2404,2405..2416)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2450..2466)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2503..2521)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2542..2558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2555..2577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2566..2582
                     /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(2590..2596,2597..2614,2615..2620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2635..2656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2773..2790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2944..3532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3024..3043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3703..4494,4495..4563))
                     /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     4326..4345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4564..4668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4831..4852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5201..5212,5213..5375)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5744..5865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5988..6698)
                     /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(6068..6091)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6266..6285)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6535..6553
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6695..6704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     7346..7362
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7382..7402
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7404..7423
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7404..7422
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7431..7538
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7447..7463
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7448..7464
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7498..7514)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7551..7567,1..2))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttcaagtttg tacaaaaaag caggctcgct acagattaaa gattggagtt ttaaactctt
       61 atgtgcattt ttatcattct attttatata ataatattta ataataataa taataataac
      121 tggctttaaa taaatacaca agggattatt tattaattta ttatgattca ttaattaatt
      181 agataaatgc aatttatgta ttccttccct ttatactatg tatgtatctt aaaatggtag
      241 tatttattat aattatttta tttaattgat taaaacattt atcaaacatg cagaactgtc
      301 cttccccaaa acagccacta gggaccggaa atgtacaaca aacattttca ggcaccagtt
      361 aacatcacct aacaccctca catgtacctg acagtactgt acataactat tgaaaagttg
      421 tctgaatatt gttcggcact attaatatac ctgcagtcgg ctacgaatga tattcctctc
      481 caacatcaac acaaagtaag ccatgaaact tcaccgtcac aattacgtgg tgaaaccgcg
      541 ttgaatattt agcaaataat gcaacttccc ggaaatacat tgatgggctg catttgtttt
      601 aagcagagag gaggcggagt tcagcgtttc cggagagcgc gcgcctgaag cagcgcatcg
      661 cggccagttg atcgccgctc ggacactcaa gctccgccac cctgcctcct ccagcgacat
      721 tatcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      781 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      841 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      901 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      961 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1021 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1081 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1141 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1201 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1261 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1321 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1381 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1441 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     1501 gggcccggga tccaccggat ctagataact gatacccagc tttcttgtac aaagtggggc
     1561 gcgcctacgt aactagtctc gagcaacttt attatacata gttgataatt cactggccgt
     1621 cgttttacgg taccatcgat gatgatccag acatgataag atacattgat gagtttggac
     1681 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1741 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1801 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1861 aatgtggtat ggctgattat gatcctctag atcagatctg cgaagatacg gccacgggtg
     1921 ctcttgatcc tgtggctgat tttggactgt gctgctcgca gctgctgatg aatcacatac
     1981 ttcctccatt ttcttccact gattgactgt tataatttcc ctaatttcca ggtcaaggtg
     2041 ctgtgcattg tggtaataga tgtgacatga cgtcacttcc aaaggaccaa tgaacatgtc
     2101 tgaccaattt catataatgt gaaaacgatt ttcataggca gaataaataa catttaaatt
     2161 aaactgggca tcagcgcaat tcaattggtt tggtaatagc aagggaaaat agaatgaagt
     2221 gatctccaaa aaataagtac tttttgactg taaataaaat tgtaaggagt aaaaagtact
     2281 tttttttcta aaaaaatgta attaagtaaa agtaaaagta ttgattttta attgtactca
     2341 agtaaagtaa aaatccccaa aaataatact taagtacagt aatcaagtaa aattactcaa
     2401 gtactttaca cctctggttc ttgaccccct accttcagca agcccagcag atccactagt
     2461 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     2521 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aaggcatttt
     3601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4681 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4741 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4801 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4861 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccacgctgg
     4921 ctaagaactc atcagcctcc ccggtccatc tacccacgta ccaatgcacc aattggccac
     4981 aatgacggct actacatggt gccattcctt cctctttata ggaatggaga ctacctcctg
     5041 tccaacaatg ctcttggata cgagtacgcc tacctgttgg acccaggtca ttgcacaaca
     5101 ccagaaatgc cctctgatct gcaaaagacg tgaatatctg ttcagacacc catatccact
     5161 ctgttccaca caggtcagag gtttgtccag gagttcttga cagaggtgta aaaagtactc
     5221 aaaaatttta ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta
     5281 aaagtatctg gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta
     5341 cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa actagagatt cttgtttaag
     5401 cttttaatct caaaaaacat taaatgaaat gcatacaagg ttttatcctg ctttagaact
     5461 gtttgtattt aattatcaaa ctataagaca gacaatctaa tgccagtaca cgctactcaa
     5521 agttgtaaaa cctcagattt aacttcagta gaagctgatt ctcaaaattg ttagtgtcaa
     5581 gcctagctct tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc tcacaggcag
     5641 ccgatgcggg aagaggtgta ttagtcttga tagagaggct gcaaatagca ggaaacgtga
     5701 gcagagactc cctggtgtct gaaacacagg cccatttacg ccttaagata cattgatgag
     5761 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     5821 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5881 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5941 ctctacaaat gtggtatggc tgattatgat ctagagtcgc ggccgcttta cttgtacagc
     6001 tcgtccatgc cgccggtgga gtggtggccc tcggcgcgct cgtactgttc cacgatggtg
     6061 tagtcctcgt tgtgggaggt gatgtccagc ttgatgttga cgttgtaggc gccgggcagc
     6121 tgcacgggct tcttggcctt gtaggtggtc ttgacctcgg cgtcgtagtg gccgccgtcc
     6181 ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc
     6241 tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag
     6301 ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg
     6361 gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg
     6421 gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag
     6481 ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc
     6541 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc
     6601 tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg
     6661 atgatggcca tgttgtcatc ctcgcccttg ctcaccatgg tggcgaccgg taatgtcaga
     6721 cctggtaact ccttactgta acttattgca cctgatgaac aaaagatgtg aactgaacgc
     6781 aagtaggagg ctaatagagg ctagtacagt acaaagatcc ccaaatgatg tgtgccggcc
     6841 ttagtggagg cagacccctg gttatatacc tgtgctgcag aaagaggggc ttacaaatgg
     6901 atctctctgg aatggcatta tctcatggtg ggaacagtgt tgtcagcaga atcagcgggg
     6961 tggactctga cctgagaatg acagtctagc actgaatggc tcagacaata ctcttgtggc
     7021 caactatcac cggaaaataa gtgcctgcca aaacaaacca gaccagatga agctacatgg
     7081 ctgtagaacg tcagtattta atgtgttgtg gccacttggt tcaccagatt taagagctaa
     7141 tttttttaaa tatcacagat aatcaatcgc tttttttttt cttgactgaa aagcttagac
     7201 tggaaaattc tttgaaaaag tttgcactac agaaaataaa tacataacat cctgcactga
     7261 atgcacacta ttaatagaaa agagagcatt attgtaggaa atgtattgta tatgtacatt
     7321 ctgtgcattt cacaattttc tcgagcagga aacagctatg accatgatta cgccaagcta
     7381 tcaactttgt atagaaaagt tggtaatacg actcactata gggcgaattg ggtaccgggc
     7441 cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat
     7501 ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     7561 gggttaa
//
