LOCUS       sequence-178592-        3883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Mid-entry clone containing promoter sequence specific for 
            maffbb driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3883)
  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 3883)
  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: pMEmafbbP1gfp
FEATURES             Location/Qualifiers
     source          1..3883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     758..775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(934..961)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1053..1139)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1188..1210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1202..1219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1203..1219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1235..1334
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1865..1874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1871..1873,1874..1876,1877..2587)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1871..1873,1874..1876,1877..2587)
                     /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(1916..1937)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2177..2196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2524..2545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2588..2653
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2674..2773)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2790..2809)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2791..2809)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2814..2830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2814..2830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2932..2951)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2943..3752
                     /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(3010..3029)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      781 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      841 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      901 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      961 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     1021 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     1081 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     1141 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     1201 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
     1261 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
     1321 caaaaaagca ggctcgctat taattttatg tgaaaataca aggaaataaa ctttcaccca
     1381 aactaatcct gtaatgcttg tcactgaaga aataaaatct taaatggcct tatagagttg
     1441 gtcagttatc agcatttgtt cattttttat tcttaaaaac gtacacatta ggcttgataa
     1501 aacaaattat atattttttt aaatattgac atacttaatt taacattatt attattatcg
     1561 gatatcagaa acggggaaac tttgacgggt tttctttaga gttttagatt tgtacatttg
     1621 aagtgtgttt tggtgcgact gacagctgtg atagggcgga gggcgtggcc tcactgcgcg
     1681 gctgctttta aagcagcgtg atttactttc ttttcaacat acatcctgcg cttgtcaccg
     1741 agagaccacc cgaagctaaa aagcgacgac aaacaggcta atagtttaag agggaggttg
     1801 gaagagcaca tccagcttga ttcagaactg gaggaattgt gttttccccc ctgaacttta
     1861 attcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1921 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1981 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2041 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2101 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2161 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2221 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2281 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2341 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2401 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2461 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2521 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2581 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     2641 gggcccggga tccaccggat ctagataact gatacccagc tttcttgtac aaagttggca
     2701 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2761 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2821 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2881 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2941 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     3001 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3061 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3121 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3181 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3241 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3301 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3361 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3421 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3481 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3541 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3601 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3661 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3721 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3781 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3841 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaa
//
