LOCUS       sequence-178530-        4005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Mid-entry clone containing promoter sequence specific for 
            clec14a driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4005)
  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 4005)
  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: pMEclecl14aP1gfp
FEATURES             Location/Qualifiers
     source          1..4005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     492..511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     745..762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(921..948)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1040..1126)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1175..1197
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1189..1206
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1190..1206
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1222..1321
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1974..1983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1980..1982,1983..1985,1986..2696)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1980..1982,1983..1985,1986..2696)
                     /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(2025..2046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2286..2305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2633..2654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2697..2762
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2783..2882)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2899..2918)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2900..2918)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2923..2939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2923..2939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3041..3060)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3052..3861
                     /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(3119..3138)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
       61 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      781 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      841 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
      901 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
      961 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     1021 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     1081 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     1141 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     1201 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
     1261 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc
     1321 tcgctagttg acttgacata tatgcacgta atgtgttctg taaatataca ataatttatt
     1381 ttccacttaa agacccttaa aaatagggca ctgatgtaaa gtacattttt aaatggcgag
     1441 gtataaagtg gaataatttt caaaacttta tactcactac tcttgaatat ttttagaagt
     1501 gctacttttt actcatattt tgagtagcag gtacttttac ttgcactaca tatttttggg
     1561 aagtaaagat acttttactt gagcatgatt tttcagtcct ctttccacca ctggctacga
     1621 tgaaacctga aaccttcaag gagatgatgg gtgtggatgc atgaacggtt tatcagccct
     1681 atgccaagaa acacaggttt cctcgtcatt taaagctcat ccgtcactct gctgtgggag
     1741 tttggcagtg ggattgtgta ttatggaagc ttcagtttca ccaggaacag atttttgttc
     1801 atagttgcga ttgttttgct tcggcagcac caccacccaa gaggaagcga atggctgtat
     1861 caaggctgaa cagacttcct gtgcttgttg tcttgggagg acctcacagg gacggctaag
     1921 aaagccaggg ggagaaaaag cagacaatat cattttagtt tgagtctgcg atcgccacca
     1981 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2041 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2101 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2161 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2221 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2281 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2341 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2401 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2461 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2521 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2581 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2641 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     2701 gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     2761 ccaccggatc tagataactg atacccagct ttcttgtaca aagttggcat tataagaaag
     2821 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     2881 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     2941 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3001 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3061 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3121 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3181 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3241 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     3301 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     3361 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     3421 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     3481 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     3541 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     3601 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     3661 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     3721 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     3781 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     3841 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     3901 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     3961 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatct
//
