LOCUS       sequence-178876-        7747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains mafbb enhancer sequence, and mafbb specific gene promoter 
            driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7747)
  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 7747)
  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;mafbbCE1-P1Egfp
FEATURES             Location/Qualifiers
     source          1..7747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     632..648
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    668..688
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1203..1227
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1758..1767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1764..1766,1767..1769,1770..2480)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1764..1766,1767..1769,1770..2480)
                     /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(1809..1830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2070..2089)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2417..2438
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2481..2546
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2567..2591)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2617..2637)
                     /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(2645..2661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2677..2811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3250..3437,3438..3449)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3483..3499)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3483..3499)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3536..3556)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3536..3554)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3575..3591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3588..3610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3599..3615
                     /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(3623..3629,3630..3647,3648..3653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3668..3689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3806..3823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3977..4565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4057..4076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4736..5527,5528..5596))
                     /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     5359..5378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5597..5701)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5864..5885)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6234..6245,6246..6408)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6777..6898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7021..7731)
                     /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(7101..7124)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7299..7318)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7568..7586
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7728..7737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtcagacctg gtaactcctt actgtaactt attgcacctg atgaacaaaa gatgtgaact
       61 gaacgcaagt aggaggctaa tagaggctag tacagtacaa agatccccaa atgatgtgtg
      121 ccggccttag tggaggcaga cccctggtta tatacctgtg ctgcagaaag aggggcttac
      181 aaatggatct ctctggaatg gcattatctc atggtgggaa cagtgttgtc agcagaatca
      241 gcggggtgga ctctgacctg agaatgacag tctagcactg aatggctcag acaatactct
      301 tgtggccaac tatcaccgga aaataagtgc ctgccaaaac aaaccagacc agatgaagct
      361 acatggctgt agaacgtcag tatttaatgt gttgtggcca cttggttcac cagatttaag
      421 agctaatttt tttaaatatc acagataatc aatcgctttt ttttttcttg actgaaaagc
      481 ttagactgga aaattctttg aaaaagtttg cactacagaa aataaataca taacatcctg
      541 cactgaatgc acactattaa tagaaaagag agcattattg taggaaatgt attgtatatg
      601 tacattctgt gcatttcaca attttctcga gcaggaaaca gctatgacca tgattacgcc
      661 aagctatcaa ctttgtatag aaaagttgtt gtcggtcgtc cacactgtct gctataggaa
      721 cccgagcaac tccagttcag tttgaccact aaactgagtc aaaagtgtct gatctctgaa
      781 gcagaactcg gctggtcaga actggtctgt tcttgcttct caggctgttt tctagggtgg
      841 ataataccag tgtcctctgt ctctctcagc cttcacgtga acagcagcgg ccgtgtggca
      901 gtgaggatgg atggggatat atggagcttc ctctttcact tcctaaatag gggtctggtc
      961 aagtgactct ggtggagagt gaggggagat gaagtgaagg aatgcaatgc ggccgcaggc
     1021 tttgtggttt ccgttcagga tgtccacttg tttctgtgtt tcgagctctg acagcttgtg
     1081 ggggtgtttg tgattcattt gtgatttact gtttcacaag aacattcaat cagtgaaccg
     1141 actcagtttg agtaagatct gctactgctt ttaatatgat gctatgaggt gatgtagtgc
     1201 tgccaagttt gtacaaaaaa gcaggctcgc tattaatttt atgtgaaaat acaaggaaat
     1261 aaactttcac ccaaactaat cctgtaatgc ttgtcactga agaaataaaa tcttaaatgg
     1321 ccttatagag ttggtcagtt atcagcattt gttcattttt tattcttaaa aacgtacaca
     1381 ttaggcttga taaaacaaat tatatatttt tttaaatatt gacatactta atttaacatt
     1441 attattatta tcggatatca gaaacgggga aactttgacg ggttttcttt agagttttag
     1501 atttgtacat ttgaagtgtg ttttggtgcg actgacagct gtgatagggc ggagggcgtg
     1561 gcctcactgc gcggctgctt ttaaagcagc gtgatttact ttcttttcaa catacatcct
     1621 gcgcttgtca ccgagagacc acccgaagct aaaaagcgac gacaaacagg ctaatagttt
     1681 aagagggagg ttggaagagc acatccagct tgattcagaa ctggaggaat tgtgttttcc
     1741 cccctgaact ttaattcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1801 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1861 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1921 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1981 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2041 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2101 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2161 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2221 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2281 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2341 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2401 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2461 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag
     2521 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgataccc agctttcttg
     2581 tacaaagtgg ggcgcgccta cgtaactagt ctcgagcaac tttattatac atagttgata
     2641 attcactggc cgtcgtttta cggtaccatc gatgatgatc cagacatgat aagatacatt
     2701 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2761 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2821 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2881 taaaacctct acaaatgtgg tatggctgat tatgatcctc tagatcagat ctgcgaagat
     2941 acggccacgg gtgctcttga tcctgtggct gattttggac tgtgctgctc gcagctgctg
     3001 atgaatcaca tacttcctcc attttcttcc actgattgac tgttataatt tccctaattt
     3061 ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca tgacgtcact tccaaaggac
     3121 caatgaacat gtctgaccaa tttcatataa tgtgaaaacg attttcatag gcagaataaa
     3181 taacatttaa attaaactgg gcatcagcgc aattcaattg gtttggtaat agcaagggaa
     3241 aatagaatga agtgatctcc aaaaaataag tactttttga ctgtaaataa aattgtaagg
     3301 agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt
     3361 ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac agtaatcaag
     3421 taaaattact caagtacttt acacctctgg ttcttgaccc cctaccttca gcaagcccag
     3481 cagatccact agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     3541 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3601 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3661 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3721 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3781 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3841 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3901 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3961 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4021 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4081 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4141 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4201 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4261 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4321 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4381 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4441 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4501 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4561 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4621 gttaaggcat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4681 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4741 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4801 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4861 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4921 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4981 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5041 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5101 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5161 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5221 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5281 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5341 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5401 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5461 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5521 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5581 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5641 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5701 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5761 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5821 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5881 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5941 tggcccacgc tggctaagaa ctcatcagcc tccccggtcc atctacccac gtaccaatgc
     6001 accaattggc cacaatgacg gctactacat ggtgccattc cttcctcttt ataggaatgg
     6061 agactacctc ctgtccaaca atgctcttgg atacgagtac gcctacctgt tggacccagg
     6121 tcattgcaca acaccagaaa tgccctctga tctgcaaaag acgtgaatat ctgttcagac
     6181 acccatatcc actctgttcc acacaggtca gaggtttgtc caggagttct tgacagaggt
     6241 gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa gtacttaggg aaaattttac
     6301 tcaattaaaa gtaaaagtat ctggctagaa tcttacttga gtaaaagtaa aaaagtactc
     6361 cattaaaatt gtacttgagt attaaggaag taaaagtaaa agcaagaaag aaaactagag
     6421 attcttgttt aagcttttaa tctcaaaaaa cattaaatga aatgcataca aggttttatc
     6481 ctgctttaga actgtttgta tttaattatc aaactataag acagacaatc taatgccagt
     6541 acacgctact caaagttgta aaacctcaga tttaacttca gtagaagctg attctcaaaa
     6601 ttgttagtgt caagcctagc tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc
     6661 ctctcacagg cagccgatgc gggaagaggt gtattagtct tgatagagag gctgcaaata
     6721 gcaggaaacg tgagcagaga ctccctggtg tctgaaacac aggcccattt acgccttaag
     6781 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6841 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     6901 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     6961 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct
     7021 ttacttgtac agctcgtcca tgccgccggt ggagtggtgg ccctcggcgc gctcgtactg
     7081 ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt tgacgttgta
     7141 ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cggcgtcgta
     7201 gtggccgccg tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc
     7261 ggggtacatc cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc
     7321 gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag
     7381 ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca
     7441 cttgaagccc tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac
     7501 gtaggccttg gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg
     7561 gccgcccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc
     7621 ctcgccctcg atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg
     7681 catgaactcc ttgatgatgg ccatgttgtc atcctcgccc ttgctcacca tggtggcgac
     7741 cggtaat
//
