LOCUS       sequence-178595-        4103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Mid-entry clone containing promoter sequence specific for 
            She driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4103)
  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 4103)
  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: pMEsheP1gfp
FEATURES             Location/Qualifiers
     source          1..4103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     751..768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(927..954)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1046..1132)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1181..1203
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1195..1212
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1196..1212
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1228..1327
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2078..2087
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2084..2086,2087..2089,2090..2800)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2084..2086,2087..2089,2090..2800)
                     /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(2129..2150)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2390..2409)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2737..2758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2801..2866
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2887..2986)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3003..3022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3004..3022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3027..3043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3027..3043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3145..3164)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3156..3965
                     /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(3223..3242)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      841 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      901 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      961 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     1021 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     1081 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     1141 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     1201 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
     1261 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
     1321 gcaggctcgc taaccacttg aaaattctaa tcttttttcc atattttaat taaaaagctg
     1381 tcatatgttt ttattatatc actgaaaatc agggttgttt ctaaacacct attcttaact
     1441 gttctgaagt ttaaactgca tggtctaaaa atgaacagaa acatcgccct ttttttctgt
     1501 cttatatggt tgtacgtgtg tacctgcgtg cgcgtgtgtt ctttaatatg taattttttg
     1561 tttatgtgtg tttatttttc aaagtcttgc acaaacttct tagcactctt ttcccctgaa
     1621 actactagct gatgtttttt aatcacataa gaatttattt agcccacata taattgtttt
     1681 gctctttatt gttagcactt aaagtgcttt ataaattaaa taaaattgaa tcattttttg
     1741 cgctgattta acttaatcat aatcaattaa actaaagggt tcaaatgcgc ctcaacaggt
     1801 gtccagcgaa taagaaagtt gccattgagg ggaacagaag ctgcagaggt gcagaaacca
     1861 cccccacccg cagctgtagc tgcagcacaa aagcacgtac atccacatcc ccttcacttc
     1921 atctagactg tcgccaggag tggatatacc gggaatggga tcttcagcga gagaaaagaa
     1981 aaacttcaaa ttcttccgcg tttatgccct tcagaggctc ctctttatct tgcgggaata
     2041 attgggagca aactaaaaca agacaataca ttccatcgcc accatggtga gcaagggcga
     2101 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2161 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2221 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2281 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2341 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2401 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2461 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2521 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2581 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2641 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2701 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2761 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     2821 tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata
     2881 actgataccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     2941 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     3001 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     3061 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3121 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3181 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3241 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     3301 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3361 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3421 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     3481 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3541 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     3601 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     3661 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     3721 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     3781 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     3841 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     3901 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     3961 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     4021 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     4081 cagaccccgt agaaaagatc aaa
//
