LOCUS       sequence-178951-        7445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains dusp5 specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7445)
  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 7445)
  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;dusp5P1Egfp
FEATURES             Location/Qualifiers
     source          1..7445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..31
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(614..616,617..619,620..1330)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(614..616,617..619,620..1330)
                     /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(659..680)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(920..939)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1267..1288
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1331..1396
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1417..1441)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1467..1487)
                     /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(1495..1511)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1527..1661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2100..2287,2288..2299)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2333..2349)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2386..2404)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2425..2441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2438..2460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2449..2465
                     /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(2473..2479,2480..2497,2498..2503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2518..2539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2656..2673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2827..3415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2907..2926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3586..4377,4378..4446))
                     /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     4209..4228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4447..4551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4714..4735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5084..5095,5096..5258)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5627..5748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5871..6581)
                     /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(5951..5974)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6149..6168)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6418..6436
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6578..6587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     7229..7245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7265..7285
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7287..7306
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7287..7305
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7314..7421
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7330..7346
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7331..7347
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7381..7397)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7434..7445,1..7))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gttaattcaa gtttgtacaa aaaagcaggc tcgctattca gtcaaatttt gccaacaact
       61 tgtttttatg aaatacaaag gaccagattc agctttaatt ttcttactga agaaaaaagt
      121 tgtttatata tgatacatta aaagcatatt tttgtttttg aggtatttct tttaagtgct
      181 tgaactttta aagtgacaaa ctgaagtacc tcatcatcct ttgttggtgt gggcgataaa
      241 atagcctagt tgtcctaaaa taaccttcta tgttgtcgtt ttcaaatact ttcaaggtta
      301 ttacaataaa tgagcgcttg aattacaaga aagtgacaca tatgagcagc accgtgatga
      361 accgatgagt taaatgttca gtatttattc gcctcactcg gttttcaagg gcgggttcag
      421 tcaacagaat gtgggcgcgc tcccgtgtcg gtgtgtccat atatggttcg tgacgtcaag
      481 cgccggaggt tccattcaca taaaaccgat cagacggaat ccccttggag gaaatgtaga
      541 ctcaggctca gtacgcaggg tttagtaggc aacctacacc cctgaaacat ccacatccct
      601 atttatcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      661 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      721 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      781 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      841 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      901 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      961 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1021 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1081 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1141 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1201 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1261 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1321 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
     1381 cgcgggcccg ggatccaccg gatctagata actgataccc agctttcttg tacaaagtgg
     1441 ggcgcgccta cgtaactagt ctcgagcaac tttattatac atagttgata attcactggc
     1501 cgtcgtttta cggtaccatc gatgatgatc cagacatgat aagatacatt gatgagtttg
     1561 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1621 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1681 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1741 acaaatgtgg tatggctgat tatgatcctc tagatcagat ctgcgaagat acggccacgg
     1801 gtgctcttga tcctgtggct gattttggac tgtgctgctc gcagctgctg atgaatcaca
     1861 tacttcctcc attttcttcc actgattgac tgttataatt tccctaattt ccaggtcaag
     1921 gtgctgtgca ttgtggtaat agatgtgaca tgacgtcact tccaaaggac caatgaacat
     1981 gtctgaccaa tttcatataa tgtgaaaacg attttcatag gcagaataaa taacatttaa
     2041 attaaactgg gcatcagcgc aattcaattg gtttggtaat agcaagggaa aatagaatga
     2101 agtgatctcc aaaaaataag tactttttga ctgtaaataa aattgtaagg agtaaaaagt
     2161 actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt ttaattgtac
     2221 tcaagtaaag taaaaatccc caaaaataat acttaagtac agtaatcaag taaaattact
     2281 caagtacttt acacctctgg ttcttgaccc cctaccttca gcaagcccag cagatccact
     2341 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     2401 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2461 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2521 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaaggcat
     3481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4561 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4621 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4681 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4741 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc
     4801 tggctaagaa ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc
     4861 cacaatgacg gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc
     4921 ctgtccaaca atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca
     4981 acaccagaaa tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc
     5041 actctgttcc acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta
     5101 ctcaaaaatt ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa
     5161 gtaaaagtat ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt
     5221 gtacttgagt attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt
     5281 aagcttttaa tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga
     5341 actgtttgta tttaattatc aaactataag acagacaatc taatgccagt acacgctact
     5401 caaagttgta aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt
     5461 caagcctagc tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg
     5521 cagccgatgc gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg
     5581 tgagcagaga ctccctggtg tctgaaacac aggcccattt acgccttaag atacattgat
     5641 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5701 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     5761 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     5821 aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct ttacttgtac
     5881 agctcgtcca tgccgccggt ggagtggtgg ccctcggcgc gctcgtactg ttccacgatg
     5941 gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt tgacgttgta ggcgccgggc
     6001 agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cggcgtcgta gtggccgccg
     6061 tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc
     6121 cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg
     6181 aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc
     6241 tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
     6301 tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg
     6361 gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg
     6421 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
     6481 atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc
     6541 ttgatgatgg ccatgttgtc atcctcgccc ttgctcacca tggtggcgac cggtaatgtc
     6601 agacctggta actccttact gtaacttatt gcacctgatg aacaaaagat gtgaactgaa
     6661 cgcaagtagg aggctaatag aggctagtac agtacaaaga tccccaaatg atgtgtgccg
     6721 gccttagtgg aggcagaccc ctggttatat acctgtgctg cagaaagagg ggcttacaaa
     6781 tggatctctc tggaatggca ttatctcatg gtgggaacag tgttgtcagc agaatcagcg
     6841 gggtggactc tgacctgaga atgacagtct agcactgaat ggctcagaca atactcttgt
     6901 ggccaactat caccggaaaa taagtgcctg ccaaaacaaa ccagaccaga tgaagctaca
     6961 tggctgtaga acgtcagtat ttaatgtgtt gtggccactt ggttcaccag atttaagagc
     7021 taattttttt aaatatcaca gataatcaat cgcttttttt tttcttgact gaaaagctta
     7081 gactggaaaa ttctttgaaa aagtttgcac tacagaaaat aaatacataa catcctgcac
     7141 tgaatgcaca ctattaatag aaaagagagc attattgtag gaaatgtatt gtatatgtac
     7201 attctgtgca tttcacaatt ttctcgagca ggaaacagct atgaccatga ttacgccaag
     7261 ctatcaactt tgtatagaaa agttggtaat acgactcact atagggcgaa ttgggtaccg
     7321 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     7381 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     7441 tgagg
//
