LOCUS       sequence-178752-        7772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains dusp5 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 7772)
  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 7772)
  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;dusp5CE1-basEgfp
FEATURES             Location/Qualifiers
     source          1..7772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     639..655
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    675..695
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1458..1482
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1559..1568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1565..1567,1568..1570,1571..2281)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1565..1567,1568..1570,1571..2281)
                     /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(1610..1631)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1871..1890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2218..2239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2282..2347
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2471..2592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2599..2623)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2649..2669)
                     /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(2677..2693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2709..2843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3282..3469,3470..3481)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3515..3531)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3515..3531)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3568..3588)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3568..3586)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3607..3623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3620..3642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3631..3647
                     /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(3655..3661,3662..3679,3680..3685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3700..3721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3838..3855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4009..4597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4089..4108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4768..5559,5560..5628))
                     /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     5391..5410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5629..5733)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5896..5917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6266..6277,6278..6440)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6809..6930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7053..7763)
                     /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(7133..7156)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7331..7350)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7600..7618
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7760..7769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cggtaatgtc agacctggta actccttact gtaacttatt gcacctgatg aacaaaagat
       61 gtgaactgaa cgcaagtagg aggctaatag aggctagtac agtacaaaga tccccaaatg
      121 atgtgtgccg gccttagtgg aggcagaccc ctggttatat acctgtgctg cagaaagagg
      181 ggcttacaaa tggatctctc tggaatggca ttatctcatg gtgggaacag tgttgtcagc
      241 agaatcagcg gggtggactc tgacctgaga atgacagtct agcactgaat ggctcagaca
      301 atactcttgt ggccaactat caccggaaaa taagtgcctg ccaaaacaaa ccagaccaga
      361 tgaagctaca tggctgtaga acgtcagtat ttaatgtgtt gtggccactt ggttcaccag
      421 atttaagagc taattttttt aaatatcaca gataatcaat cgcttttttt tttcttgact
      481 gaaaagctta gactggaaaa ttctttgaaa aagtttgcac tacagaaaat aaatacataa
      541 catcctgcac tgaatgcaca ctattaatag aaaagagagc attattgtag gaaatgtatt
      601 gtatatgtac attctgtgca tttcacaatt ttctcgagca ggaaacagct atgaccatga
      661 ttacgccaag ctatcaactt tgtatagaaa agttggttaa aacaactaca ctttcataaa
      721 ggtcaacatc tacatatcta caagcattac attttttgtt ttcaactgct gtataagcaa
      781 catgtgctaa catgtaatag gcggcaagac agctgagtaa aatctgcaaa acacacacac
      841 acacacacac tcatgcactt gagcagcaga tgaagatctg tctgtggttc attagtaatg
      901 aggctctcgc agacacacct catctccacc ctgctgctgt ggaactggga gttttacgcc
      961 acaccacagt ttcattctca ctctaaatgg aaactgtgcg tgcactcccc aatttaagca
     1021 cacaacctaa tttttcaaaa gatggccaca aagtttgagc tgaccagttg gatgttgtgt
     1081 tgtttttgct gagtcctcga ggtatgggat gtttacatgc cctgaggatg tgcattgttt
     1141 ctcatgcctg gtcttgtagc tgccagtcta gatctgctga gagctgttcc catttcctgc
     1201 tttacaggaa gtgtcggccc ccactggtct ttgtcactca gagagacagg aagaggaaac
     1261 ggcccacagg aaacatctcc cccacttgct acgaatcatc ccgcaacatg agcctacaag
     1321 cacaaactgt tttcatttag ttagtttatg gtagaacaca ctgtgaaagt gaagatgttt
     1381 gcaatggtct gacctaaatg cttttatttg tgtgacacaa tgtaattagg acagttaatg
     1441 catgattaag tagtcgaaca agtttgtaca aaaaagcagg ctagactcta gagggtatat
     1501 aatggatctc gcgtctcagc ctcactttga gctcctccac acgctagcgc taccggtcgc
     1561 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1621 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1681 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1741 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1801 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1861 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1921 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1981 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2041 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2101 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2161 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2221 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2281 gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc
     2341 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2401 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2461 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2521 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2581 caatgtatct taacgcgtac ccagctttct tgtacaaagt ggggcgcgcc tacgtaacta
     2641 gtctcgagca actttattat acatagttga taattcactg gccgtcgttt tacggtacca
     2701 tcgatgatga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2761 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2821 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2881 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     2941 attatgatcc tctagatcag atctgcgaag atacggccac gggtgctctt gatcctgtgg
     3001 ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct ccattttctt
     3061 ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg cattgtggta
     3121 atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc aatttcatat
     3181 aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact gggcatcagc
     3241 gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct ccaaaaaata
     3301 agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa
     3361 atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa agtaaaaatc
     3421 cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact ttacacctct
     3481 ggttcttgac cccctacctt cagcaagccc agcagatcca ctagttctag agcggccgcc
     3541 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     3601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4621 ggggtctgac gctcagtgga acgaaaactc acgttaaggc attttggtca tgagattatc
     4681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     5761 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5821 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5881 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5941 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac gctggctaag aactcatcag
     6001 cctccccggt ccatctaccc acgtaccaat gcaccaattg gccacaatga cggctactac
     6061 atggtgccat tccttcctct ttataggaat ggagactacc tcctgtccaa caatgctctt
     6121 ggatacgagt acgcctacct gttggaccca ggtcattgca caacaccaga aatgccctct
     6181 gatctgcaaa agacgtgaat atctgttcag acacccatat ccactctgtt ccacacaggt
     6241 cagaggtttg tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa
     6301 gtgaaagtac aagtacttag ggaaaatttt actcaattaa aagtaaaagt atctggctag
     6361 aatcttactt gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga gtattaagga
     6421 agtaaaagta aaagcaagaa agaaaactag agattcttgt ttaagctttt aatctcaaaa
     6481 aacattaaat gaaatgcata caaggtttta tcctgcttta gaactgtttg tatttaatta
     6541 tcaaactata agacagacaa tctaatgcca gtacacgcta ctcaaagttg taaaacctca
     6601 gatttaactt cagtagaagc tgattctcaa aattgttagt gtcaagccta gctcttttgg
     6661 ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag
     6721 gtgtattagt cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg
     6781 tgtctgaaac acaggcccat ttacgcctta agatacattg atgagtttgg acaaaccaca
     6841 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6901 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     6961 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     7021 atggctgatt atgatctaga gtcgcggccg ctttacttgt acagctcgtc catgccgccg
     7081 gtggagtggt ggccctcggc gcgctcgtac tgttccacga tggtgtagtc ctcgttgtgg
     7141 gaggtgatgt ccagcttgat gttgacgttg taggcgccgg gcagctgcac gggcttcttg
     7201 gccttgtagg tggtcttgac ctcggcgtcg tagtggccgc cgtccttcag cttcagcctc
     7261 tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc
     7321 cagcccatgg tcttcttctg cattacgggg ccgtcggagg ggaagttggt gccgcgcagc
     7381 ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac ggtcaccacg
     7441 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     7501 aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta catgaactga
     7561 ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctt cagcttggcg
     7621 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     7681 ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgttg
     7741 tcatcctcgc ccttgctcac catggtggcg ac
//
