LOCUS       sequence-190346-        7399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains mafbb specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7399)
  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 7399)
  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: pT2-cryR;mafbbP1Egfp
FEATURES             Location/Qualifiers
     source          1..7399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(510..521,522..684)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    1053..1174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1297..2007)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGHHSTGGMDELYK"
     primer_bind     complement(1377..1400)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1575..1594)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1844..1862
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2004..2013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2655..2671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2691..2711
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     2713..2732
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2713..2731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2740..2847
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2756..2772
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2757..2773
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2807..2823)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2860..2878)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2878..2902
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3433..3442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3439..3441,3442..3444,3445..4155)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3439..3441,3442..3444,3445..4155)
                     /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(3484..3505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3745..3764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4092..4113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4156..4221
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(4242..4266)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4292..4312)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(4320..4336)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4352..4486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(4925..5112,5113..5124)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5158..5174)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5211..5229)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5250..5266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5263..5285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5274..5290
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5298..5304,5305..5322,5323..5328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5343..5364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5481..5498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5652..6240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5732..5751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6411..7202,7203..7271))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7034..7053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7272..7376)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccacgctggc taagaactca
      241 tcagcctccc cggtccatct acccacgtac caatgcacca attggccaca atgacggcta
      301 ctacatggtg ccattccttc ctctttatag gaatggagac tacctcctgt ccaacaatgc
      361 tcttggatac gagtacgcct acctgttgga cccaggtcat tgcacaacac cagaaatgcc
      421 ctctgatctg caaaagacgt gaatatctgt tcagacaccc atatccactc tgttccacac
      481 aggtcagagg tttgtccagg agttcttgac agaggtgtaa aaagtactca aaaattttac
      541 tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg
      601 ctagaatctt acttgagtaa aagtaaaaaa gtactccatt aaaattgtac ttgagtatta
      661 aggaagtaaa agtaaaagca agaaagaaaa ctagagattc ttgtttaagc ttttaatctc
      721 aaaaaacatt aaatgaaatg catacaaggt tttatcctgc tttagaactg tttgtattta
      781 attatcaaac tataagacag acaatctaat gccagtacac gctactcaaa gttgtaaaac
      841 ctcagattta acttcagtag aagctgattc tcaaaattgt tagtgtcaag cctagctctt
      901 ttggggctga aaagcaatcc tgcagtgctg aaaagcctct cacaggcagc cgatgcggga
      961 agaggtgtat tagtcttgat agagaggctg caaatagcag gaaacgtgag cagagactcc
     1021 ctggtgtctg aaacacaggc ccatttacgc cttaagatac attgatgagt ttggacaaac
     1081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1141 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1201 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1261 tggtatggct gattatgatc tagagtcgcg gccgctttac ttgtacagct cgtccatgcc
     1321 gccggtggag tggtggccct cggcgcgctc gtactgttcc acgatggtgt agtcctcgtt
     1381 gtgggaggtg atgtccagct tgatgttgac gttgtaggcg ccgggcagct gcacgggctt
     1441 cttggccttg taggtggtct tgacctcggc gtcgtagtgg ccgccgtcct tcagcttcag
     1501 cctctgcttg atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc
     1561 ctcccagccc atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg
     1621 cagcttcacc ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac
     1681 cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag
     1741 cttcaagtag tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgaa
     1801 ctgaggggac aggatgtccc aggcgaaggg cagggggccg cccttggtca ccttcagctt
     1861 ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg
     1921 gccgttcacg gagccctcca tgtgcacctt gaagcgcatg aactccttga tgatggccat
     1981 gttgtcatcc tcgcccttgc tcaccatggt ggcgaccggt aatgtcagac ctggtaactc
     2041 cttactgtaa cttattgcac ctgatgaaca aaagatgtga actgaacgca agtaggaggc
     2101 taatagaggc tagtacagta caaagatccc caaatgatgt gtgccggcct tagtggaggc
     2161 agacccctgg ttatatacct gtgctgcaga aagaggggct tacaaatgga tctctctgga
     2221 atggcattat ctcatggtgg gaacagtgtt gtcagcagaa tcagcggggt ggactctgac
     2281 ctgagaatga cagtctagca ctgaatggct cagacaatac tcttgtggcc aactatcacc
     2341 ggaaaataag tgcctgccaa aacaaaccag accagatgaa gctacatggc tgtagaacgt
     2401 cagtatttaa tgtgttgtgg ccacttggtt caccagattt aagagctaat ttttttaaat
     2461 atcacagata atcaatcgct tttttttttc ttgactgaaa agcttagact ggaaaattct
     2521 ttgaaaaagt ttgcactaca gaaaataaat acataacatc ctgcactgaa tgcacactat
     2581 taatagaaaa gagagcatta ttgtaggaaa tgtattgtat atgtacattc tgtgcatttc
     2641 acaattttct cgagcaggaa acagctatga ccatgattac gccaagctat caactttgta
     2701 tagaaaagtt ggtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag
     2761 gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc cactagttct
     2821 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattca
     2881 agtttgtaca aaaaagcagg ctcgctatta attttatgtg aaaatacaag gaaataaact
     2941 ttcacccaaa ctaatcctgt aatgcttgtc actgaagaaa taaaatctta aatggcctta
     3001 tagagttggt cagttatcag catttgttca ttttttattc ttaaaaacgt acacattagg
     3061 cttgataaaa caaattatat atttttttaa atattgacat acttaattta acattattat
     3121 tattatcgga tatcagaaac ggggaaactt tgacgggttt tctttagagt tttagatttg
     3181 tacatttgaa gtgtgttttg gtgcgactga cagctgtgat agggcggagg gcgtggcctc
     3241 actgcgcggc tgcttttaaa gcagcgtgat ttactttctt ttcaacatac atcctgcgct
     3301 tgtcaccgag agaccacccg aagctaaaaa gcgacgacaa acaggctaat agtttaagag
     3361 ggaggttgga agagcacatc cagcttgatt cagaactgga ggaattgtgt tttcccccct
     3421 gaactttaat tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3481 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3541 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3601 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3661 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3721 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3781 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3841 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3901 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3961 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4021 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     4081 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4141 gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac
     4201 ggtaccgcgg gcccgggatc caccggatct agataactga tacccagctt tcttgtacaa
     4261 agtggggcgc gcctacgtaa ctagtctcga gcaactttat tatacatagt tgataattca
     4321 ctggccgtcg ttttacggta ccatcgatga tgatccagac atgataagat acattgatga
     4381 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4441 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4501 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4561 cctctacaaa tgtggtatgg ctgattatga tcctctagat cagatctgcg aagatacggc
     4621 cacgggtgct cttgatcctg tggctgattt tggactgtgc tgctcgcagc tgctgatgaa
     4681 tcacatactt cctccatttt cttccactga ttgactgtta taatttccct aatttccagg
     4741 tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg tcacttccaa aggaccaatg
     4801 aacatgtctg accaatttca tataatgtga aaacgatttt cataggcaga ataaataaca
     4861 tttaaattaa actgggcatc agcgcaattc aattggtttg gtaatagcaa gggaaaatag
     4921 aatgaagtga tctccaaaaa ataagtactt tttgactgta aataaaattg taaggagtaa
     4981 aaagtacttt tttttctaaa aaaatgtaat taagtaaaag taaaagtatt gatttttaat
     5041 tgtactcaag taaagtaaaa atccccaaaa ataatactta agtacagtaa tcaagtaaaa
     5101 ttactcaagt actttacacc tctggttctt gaccccctac cttcagcaag cccagcagat
     5161 ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     5221 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5281 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5341 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5401 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5461 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6301 ggcattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7381 tttccccgaa aagtgccac
//
