LOCUS       sequence-178705-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains mafbb enhancer sequence and a basal promoter driving 
            expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7524)
  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 7524)
  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-basEgfp
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     621..637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    657..677
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1192..1216
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1293..1302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1299..1301,1302..1304,1305..2015)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1299..1301,1302..1304,1305..2015)
                     /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(1344..1365)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1605..1624)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1952..1973
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2016..2081
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2205..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2333..2357)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2383..2403)
                     /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(2411..2427)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2443..2577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3016..3203,3204..3215)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3249..3265)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3249..3265)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3302..3322)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3302..3320)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3341..3357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3354..3376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3365..3381
                     /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(3389..3395,3396..3413,3414..3419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3434..3455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3572..3589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3743..4331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3823..3842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4502..5293,5294..5362))
                     /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     5125..5144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5363..5467)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5630..5651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6000..6011,6012..6174)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6543..6664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6787..7497)
                     /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(6867..6890)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7065..7084)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7334..7352
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7494..7503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 taactcctta ctgtaactta ttgcacctga tgaacaaaag atgtgaactg aacgcaagta
       61 ggaggctaat agaggctagt acagtacaaa gatccccaaa tgatgtgtgc cggccttagt
      121 ggaggcagac ccctggttat atacctgtgc tgcagaaaga ggggcttaca aatggatctc
      181 tctggaatgg cattatctca tggtgggaac agtgttgtca gcagaatcag cggggtggac
      241 tctgacctga gaatgacagt ctagcactga atggctcaga caatactctt gtggccaact
      301 atcaccggaa aataagtgcc tgccaaaaca aaccagacca gatgaagcta catggctgta
      361 gaacgtcagt atttaatgtg ttgtggccac ttggttcacc agatttaaga gctaattttt
      421 ttaaatatca cagataatca atcgcttttt tttttcttga ctgaaaagct tagactggaa
      481 aattctttga aaaagtttgc actacagaaa ataaatacat aacatcctgc actgaatgca
      541 cactattaat agaaaagaga gcattattgt aggaaatgta ttgtatatgt acattctgtg
      601 catttcacaa ttttctcgag caggaaacag ctatgaccat gattacgcca agctatcaac
      661 tttgtataga aaagttgttg tcggtcgtcc acactgtctg ctataggaac ccgagcaact
      721 ccagttcagt ttgaccacta aactgagtca aaagtgtctg atctctgaag cagaactcgg
      781 ctggtcagaa ctggtctgtt cttgcttctc aggctgtttt ctagggtgga taataccagt
      841 gtcctctgtc tctctcagcc ttcacgtgaa cagcagcggc cgtgtggcag tgaggatgga
      901 tggggatata tggagcttcc tctttcactt cctaaatagg ggtctggtca agtgactctg
      961 gtggagagtg aggggagatg aagtgaagga atgcaatgcg gccgcaggct ttgtggtttc
     1021 cgttcaggat gtccacttgt ttctgtgttt cgagctctga cagcttgtgg gggtgtttgt
     1081 gattcatttg tgatttactg tttcacaaga acattcaatc agtgaaccga ctcagtttga
     1141 gtaagatctg ctactgcttt taatatgatg ctatgaggtg atgtagtgct gccaagtttg
     1201 tacaaaaaag caggctagac tctagagggt atataatgga tctcgcgtct cagcctcact
     1261 ttgagctcct ccacacgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1321 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1381 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1441 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1501 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1561 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1621 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1681 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1741 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1801 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1861 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1921 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1981 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag
     2041 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     2101 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2161 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtacccagct
     2341 ttcttgtaca aagtggggcg cgcctacgta actagtctcg agcaacttta ttatacatag
     2401 ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga catgataaga
     2461 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2521 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2581 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2641 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctgc
     2701 gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg ctgctcgcag
     2761 ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt ataatttccc
     2821 taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac gtcacttcca
     2881 aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt tcataggcag
     2941 aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt ggtaatagca
     3001 agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt
     3061 gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat
     3121 tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta
     3181 atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa
     3241 gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     3301 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3361 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3421 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3481 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3541 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3601 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3661 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3721 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3781 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3841 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3901 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3961 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4021 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4081 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4141 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4201 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4261 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4321 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4381 actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4441 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4501 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4561 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4621 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4681 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4741 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4801 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4861 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4921 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4981 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5041 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5101 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5161 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5221 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5281 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5341 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5401 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5461 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5521 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5581 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5641 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5701 gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct acccacgtac
     5761 caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc ctctttatag
     5821 gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct acctgttgga
     5881 cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt gaatatctgt
     5941 tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg agttcttgac
     6001 agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa
     6061 ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa
     6121 gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca agaaagaaaa
     6181 ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg catacaaggt
     6241 tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag acaatctaat
     6301 gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag aagctgattc
     6361 tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc tgcagtgctg
     6421 aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat agagaggctg
     6481 caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc ccatttacgc
     6541 cttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     6601 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6661 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     6721 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc tagagtcgcg
     6781 gccgctttac ttgtacagct cgtccatgcc gccggtggag tggtggccct cggcgcgctc
     6841 gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tgatgttgac
     6901 gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct tgacctcggc
     6961 gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct tcagggcgcc
     7021 gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct tctgcattac
     7081 ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga actcgccgtc
     7141 ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg
     7201 ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt cggcggggtg
     7261 cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc aggcgaaggg
     7321 cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc
     7381 ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca tgtgcacctt
     7441 gaagcgcatg aactccttga tgatggccat gttgtcatcc tcgcccttgc tcaccatggt
     7501 ggcgaccggt aatgtcagac ctgg
//
