LOCUS       sequence-178731-        7878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains yrk 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 7878)
  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 7878)
  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;yrkCE1-basEgfp
FEATURES             Location/Qualifiers
     source          1..7878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     629..645
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    665..685
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1554..1578
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1655..1664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1661..1663,1664..1666,1667..2377)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1661..1663,1664..1666,1667..2377)
                     /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(1706..1727)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1967..1986)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2314..2335
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2378..2443
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2567..2688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2695..2719)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2745..2765)
                     /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(2773..2789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2805..2939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3378..3565,3566..3577)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3611..3627)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3611..3627)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3664..3684)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3664..3682)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3703..3719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3716..3738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3727..3743
                     /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(3751..3757,3758..3775,3776..3781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3796..3817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3934..3951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4105..4693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4185..4204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4864..5655,5656..5724))
                     /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     5487..5506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5725..5829)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5992..6013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6362..6373,6374..6536)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6905..7026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7149..7859)
                     /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(7229..7252)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7427..7446)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7696..7714
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7856..7865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agacctggta actccttact gtaacttatt gcacctgatg aacaaaagat gtgaactgaa
       61 cgcaagtagg aggctaatag aggctagtac agtacaaaga tccccaaatg atgtgtgccg
      121 gccttagtgg aggcagaccc ctggttatat acctgtgctg cagaaagagg ggcttacaaa
      181 tggatctctc tggaatggca ttatctcatg gtgggaacag tgttgtcagc agaatcagcg
      241 gggtggactc tgacctgaga atgacagtct agcactgaat ggctcagaca atactcttgt
      301 ggccaactat caccggaaaa taagtgcctg ccaaaacaaa ccagaccaga tgaagctaca
      361 tggctgtaga acgtcagtat ttaatgtgtt gtggccactt ggttcaccag atttaagagc
      421 taattttttt aaatatcaca gataatcaat cgcttttttt tttcttgact gaaaagctta
      481 gactggaaaa ttctttgaaa aagtttgcac tacagaaaat aaatacataa catcctgcac
      541 tgaatgcaca ctattaatag aaaagagagc attattgtag gaaatgtatt gtatatgtac
      601 attctgtgca tttcacaatt ttctcgagca ggaaacagct atgaccatga ttacgccaag
      661 ctatcaactt tgtatagaaa agttgcgttc tcagtacttt aaagagccta tgacaacaac
      721 aacaacaaca acaacatcat gcatatttaa tgtcacagta tattgcagca ttgcatactg
      781 gactaaaaag catgatgagg tttgattaat taaatggaaa acagaatgga atgtctctag
      841 gactagatta ggccataggc ttcatcactt cctgttctgt gcaaggtcac ttcctgtcta
      901 ctctcccctg gcgctatggg aggaagtggg tgcggctgtc cggttaaatt gtccgaaatg
      961 caacatggtt aaactgacca aaccagagcc tgcaatgtgg attttcagct ctccgatgtt
     1021 ctgttcacta ctgacctgat gtttagtgtg taggaaacgt tggctaaatc ccacatattg
     1081 tattcaatgg gttgcttgtt tgattagcgt gaaactggca ttgattctga tgatgaatac
     1141 atttttttgg ctttagtttt atccaattag tgcatgcata ttcagagaaa gtaagtttaa
     1201 aaagcacata tatttagttt attttccgaa agcacccgtg ctgaaatact gcgttcatta
     1261 acggcctgct cgtggaatct ggctgtggag gaaattgaca tcatgtggaa aaaggaagtg
     1321 gctgtgtttg tgacacctga ctttctgttc ccagggcaca ctctgctatt gtcttccctc
     1381 tctgcaatat tatctctcag tttctctctc actgcattgt ttcacctagt tgttgttgtt
     1441 gttgtttatt tgcctgagtg acttttatta cagtttttat gagtgtgtgg gtgtattcag
     1501 catggtctca cattttcctg ttgtgataat tgctaaatct ttaatcaggg tatacaagtt
     1561 tgtacaaaaa agcaggctag actctagagg gtatataatg gatctcgcgt ctcagcctca
     1621 ctttgagctc ctccacacgc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     1681 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1741 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1801 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1861 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1921 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1981 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2041 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2101 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2161 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2221 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2281 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2341 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
     2401 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     2461 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2521 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2581 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtacccag
     2701 ctttcttgta caaagtgggg cgcgcctacg taactagtct cgagcaactt tattatacat
     2761 agttgataat tcactggccg tcgttttacg gtaccatcga tgatgatcca gacatgataa
     2821 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2881 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2941 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     3001 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcctcta gatcagatct
     3061 gcgaagatac ggccacgggt gctcttgatc ctgtggctga ttttggactg tgctgctcgc
     3121 agctgctgat gaatcacata cttcctccat tttcttccac tgattgactg ttataatttc
     3181 cctaatttcc aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg acgtcacttc
     3241 caaaggacca atgaacatgt ctgaccaatt tcatataatg tgaaaacgat tttcataggc
     3301 agaataaata acatttaaat taaactgggc atcagcgcaa ttcaattggt ttggtaatag
     3361 caagggaaaa tagaatgaag tgatctccaa aaaataagta ctttttgact gtaaataaaa
     3421 ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt
     3481 attgattttt aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag
     3541 taatcaagta aaattactca agtactttac acctctggtt cttgaccccc taccttcagc
     3601 aagcccagca gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg
     3661 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3721 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3781 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3841 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3901 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3961 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4021 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4621 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4741 aaactcacgt taaggcattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4861 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4921 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4981 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5041 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5101 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5161 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5221 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5281 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5341 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5401 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5461 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5521 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5581 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5641 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5701 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5761 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5821 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5881 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6061 cagggcgatg gcccacgctg gctaagaact catcagcctc cccggtccat ctacccacgt
     6121 accaatgcac caattggcca caatgacggc tactacatgg tgccattcct tcctctttat
     6181 aggaatggag actacctcct gtccaacaat gctcttggat acgagtacgc ctacctgttg
     6241 gacccaggtc attgcacaac accagaaatg ccctctgatc tgcaaaagac gtgaatatct
     6301 gttcagacac ccatatccac tctgttccac acaggtcaga ggtttgtcca ggagttcttg
     6361 acagaggtgt aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa
     6421 aattttactc aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa
     6481 aagtactcca ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa
     6541 aactagagat tcttgtttaa gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag
     6601 gttttatcct gctttagaac tgtttgtatt taattatcaa actataagac agacaatcta
     6661 atgccagtac acgctactca aagttgtaaa acctcagatt taacttcagt agaagctgat
     6721 tctcaaaatt gttagtgtca agcctagctc ttttggggct gaaaagcaat cctgcagtgc
     6781 tgaaaagcct ctcacaggca gccgatgcgg gaagaggtgt attagtcttg atagagaggc
     6841 tgcaaatagc aggaaacgtg agcagagact ccctggtgtc tgaaacacag gcccatttac
     6901 gccttaagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     6961 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     7021 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     7081 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tctagagtcg
     7141 cggccgcttt acttgtacag ctcgtccatg ccgccggtgg agtggtggcc ctcggcgcgc
     7201 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag cttgatgttg
     7261 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctcg
     7321 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     7381 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     7441 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     7501 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     7561 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     7621 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     7681 ggcagggggc cgcccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     7741 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     7801 ttgaagcgca tgaactcctt gatgatggcc atgttgtcat cctcgccctt gctcaccatg
     7861 gtggcgaccg gtaatgtc
//
