LOCUS       sequence-178563-        3929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Mid-entry clone containing promoter sequence specific for 
            dusp5 driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3929)
  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 3929)
  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: pMEdusp5P1gfp
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(928..955)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1047..1133)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1182..1204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1196..1213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1197..1213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1229..1328
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1905..1914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1911..1913,1914..1916,1917..2627)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1911..1913,1914..1916,1917..2627)
                     /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(1956..1977)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2217..2236)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2564..2585
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2628..2693
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2714..2813)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2830..2849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2831..2849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2854..2870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2854..2870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2972..2991)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2983..3792
                     /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(3050..3069)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      781 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      841 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      901 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      961 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     1021 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     1081 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     1141 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
     1201 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
     1261 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
     1321 agcaggctcg ctattcagtc aaattttgcc aacaacttgt ttttatgaaa tacaaaggac
     1381 cagattcagc tttaattttc ttactgaaga aaaaagttgt ttatatatga tacattaaaa
     1441 gcatattttt gtttttgagg tatttctttt aagtgcttga acttttaaag tgacaaactg
     1501 aagtacctca tcatcctttg ttggtgtggg cgataaaata gcctagttgt cctaaaataa
     1561 ccttctatgt tgtcgttttc aaatactttc aaggttatta caataaatga gcgcttgaat
     1621 tacaagaaag tgacacatat gagcagcacc gtgatgaacc gatgagttaa atgttcagta
     1681 tttattcgcc tcactcggtt ttcaagggcg ggttcagtca acagaatgtg ggcgcgctcc
     1741 cgtgtcggtg tgtccatata tggttcgtga cgtcaagcgc cggaggttcc attcacataa
     1801 aaccgatcag acggaatccc cttggaggaa atgtagactc aggctcagta cgcagggttt
     1861 agtaggcaac ctacacccct gaaacatcca catccctatt tatcgccacc atggtgagca
     1921 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1981 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2041 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2101 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2161 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2221 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2281 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2341 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2401 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2461 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2521 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2581 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat
     2641 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     2701 ctagataact gatacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2761 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2821 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     2881 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2941 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     3001 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     3061 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     3121 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     3181 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     3241 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     3301 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3361 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     3421 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3481 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3541 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3601 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3661 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3721 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3781 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     3841 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3901 tgagcgtcag accccgtaga aaagatcaa
//
