LOCUS       sequence-178606-        4051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Mid-entry clone containing promoter sequence specific for 
            yrk driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4051)
  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 4051)
  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: pMEyrkP1gfp
FEATURES             Location/Qualifiers
     source          1..4051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     746..763
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(922..949)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1041..1127)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1176..1198
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1190..1207
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1191..1207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1223..1322
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2021..2030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2027..2029,2030..2032,2033..2743)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2027..2029,2030..2032,2033..2743)
                     /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(2072..2093)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2333..2352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2680..2701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2744..2809
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2830..2929)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2946..2965)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2947..2965)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2970..2986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2970..2986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3088..3107)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3099..3908
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3166..3185)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      721 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      781 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      841 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      901 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      961 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     1021 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     1081 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     1141 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
     1201 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
     1261 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
     1321 ctcgctacag attaaagatt ggagttttaa actcttatgt gcatttttat cattctattt
     1381 tatataataa tatttaataa taataataat aataactggc tttaaataaa tacacaaggg
     1441 attatttatt aatttattat gattcattaa ttaattagat aaatgcaatt tatgtattcc
     1501 ttccctttat actatgtatg tatcttaaaa tggtagtatt tattataatt attttattta
     1561 attgattaaa acatttatca aacatgcaga actgtccttc cccaaaacag ccactaggga
     1621 ccggaaatgt acaacaaaca ttttcaggca ccagttaaca tcacctaaca ccctcacatg
     1681 tacctgacag tactgtacat aactattgaa aagttgtctg aatattgttc ggcactatta
     1741 atatacctgc agtcggctac gaatgatatt cctctccaac atcaacacaa agtaagccat
     1801 gaaacttcac cgtcacaatt acgtggtgaa accgcgttga atatttagca aataatgcaa
     1861 cttcccggaa atacattgat gggctgcatt tgttttaagc agagaggagg cggagttcag
     1921 cgtttccgga gagcgcgcgc ctgaagcagc gcatcgcggc cagttgatcg ccgctcggac
     1981 actcaagctc cgccaccctg cctcctccag cgacattatc gccaccatgg tgagcaaggg
     2041 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2101 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2161 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2221 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2281 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2341 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2401 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2461 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2521 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2581 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2641 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2701 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg
     2761 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag
     2821 ataactgata cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     2881 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     2941 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     3001 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     3061 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     3121 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     3181 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     3241 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     3301 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     3361 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     3421 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     3481 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     3541 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     3601 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     3661 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     3721 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     3781 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     3841 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     3901 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     3961 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     4021 cgtcagaccc cgtagaaaag atcaaaggat c
//
