LOCUS       sequence-178944-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains dll4 specific gene 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;dll4P1Egfp
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      683..692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(689..691,692..694,695..1405)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(689..691,692..694,695..1405)
                     /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(734..755)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(995..1014)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1342..1363
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1406..1471
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1492..1516)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1542..1562)
                     /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(1570..1586)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1602..1736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2175..2362,2363..2374)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2408..2424)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2461..2479)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2500..2516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2513..2535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2524..2540
                     /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(2548..2554,2555..2572,2573..2578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2593..2614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2731..2748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2902..3490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2982..3001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3661..4452,4453..4521))
                     /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     4284..4303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4522..4626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4789..4810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5159..5170,5171..5333)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5702..5823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5946..6656)
                     /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(6026..6049)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6224..6243)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6493..6511
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6653..6662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     7304..7320
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7340..7360
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7362..7381
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7362..7380
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7389..7496
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7405..7421
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7406..7422
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7456..7472)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7509..7524,1..3))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 attcaagttt gtacaaaaaa gcaggctcgc taggatacta aaatgcgctt tcaggctgtc
       61 ccagcttgaa acatgaagag gctctgctcc ggaacatcag gtaaggcatg accctccact
      121 gaccgtgcac gctttcagtg atgtaatata attatccgcg caattaactg gaacgaggac
      181 aaacgacaag taataggctg tgcgttgtgc gaagtttcat ccgatttagt ttttcttcag
      241 atacgcgtat tctgcactgc atggtgtgaa atggtcagta tttaatcgtg cctgctttaa
      301 atatgccaac agatgctggg cttgttaata atcactcgcg ctcaattggt catcatcatc
      361 cgcgtagtgg tgggtattaa atgtggatat cattgttgcc aagagagggg ggctgactga
      421 gcataagtgg gcggcgttta gcgatagaca agagtttata taatgagcag cgcggacaca
      481 aagccagtat aaatactgag ggacgtaaga aacaagtctt cattcacgaa ttacacgaga
      541 agcgcgagga tgcccaacag cgcaatgctg acgcacaaag tttagaagca aaacaagaca
      601 taaccattga cactaaacac atcggtgcgc tcactgtgga ataccaaaca tgctcagtgg
      661 atatcttgag acgaagagga tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      721 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      781 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      841 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
      901 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      961 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1021 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1081 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1141 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1201 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1261 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     1321 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1381 tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgaattc
     1441 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tacccagctt
     1501 tcttgtacaa agtggggcgc gcctacgtaa ctagtctcga gcaactttat tatacatagt
     1561 tgataattca ctggccgtcg ttttacggta ccatcgatga tgatccagac atgataagat
     1621 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1681 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1741 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1801 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagat cagatctgcg
     1861 aagatacggc cacgggtgct cttgatcctg tggctgattt tggactgtgc tgctcgcagc
     1921 tgctgatgaa tcacatactt cctccatttt cttccactga ttgactgtta taatttccct
     1981 aatttccagg tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg tcacttccaa
     2041 aggaccaatg aacatgtctg accaatttca tataatgtga aaacgatttt cataggcaga
     2101 ataaataaca tttaaattaa actgggcatc agcgcaattc aattggtttg gtaatagcaa
     2161 gggaaaatag aatgaagtga tctccaaaaa ataagtactt tttgactgta aataaaattg
     2221 taaggagtaa aaagtacttt tttttctaaa aaaatgtaat taagtaaaag taaaagtatt
     2281 gatttttaat tgtactcaag taaagtaaaa atccccaaaa ataatactta agtacagtaa
     2341 tcaagtaaaa ttactcaagt actttacacc tctggttctt gaccccctac cttcagcaag
     2401 cccagcagat ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc
     2461 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2521 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2581 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3541 ctcacgttaa ggcattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3961 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4261 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4621 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4681 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4741 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4801 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4861 ggcgatggcc cacgctggct aagaactcat cagcctcccc ggtccatcta cccacgtacc
     4921 aatgcaccaa ttggccacaa tgacggctac tacatggtgc cattccttcc tctttatagg
     4981 aatggagact acctcctgtc caacaatgct cttggatacg agtacgccta cctgttggac
     5041 ccaggtcatt gcacaacacc agaaatgccc tctgatctgc aaaagacgtg aatatctgtt
     5101 cagacaccca tatccactct gttccacaca ggtcagaggt ttgtccagga gttcttgaca
     5161 gaggtgtaaa aagtactcaa aaattttact caagtgaaag tacaagtact tagggaaaat
     5221 tttactcaat taaaagtaaa agtatctggc tagaatctta cttgagtaaa agtaaaaaag
     5281 tactccatta aaattgtact tgagtattaa ggaagtaaaa gtaaaagcaa gaaagaaaac
     5341 tagagattct tgtttaagct tttaatctca aaaaacatta aatgaaatgc atacaaggtt
     5401 ttatcctgct ttagaactgt ttgtatttaa ttatcaaact ataagacaga caatctaatg
     5461 ccagtacacg ctactcaaag ttgtaaaacc tcagatttaa cttcagtaga agctgattct
     5521 caaaattgtt agtgtcaagc ctagctcttt tggggctgaa aagcaatcct gcagtgctga
     5581 aaagcctctc acaggcagcc gatgcgggaa gaggtgtatt agtcttgata gagaggctgc
     5641 aaatagcagg aaacgtgagc agagactccc tggtgtctga aacacaggcc catttacgcc
     5701 ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5761 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5821 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     5881 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct agagtcgcgg
     5941 ccgctttact tgtacagctc gtccatgccg ccggtggagt ggtggccctc ggcgcgctcg
     6001 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt gatgttgacg
     6061 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcggcg
     6121 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     6181 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     6241 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     6301 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     6361 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     6421 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     6481 agggggccgc ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     6541 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     6601 aagcgcatga actccttgat gatggccatg ttgtcatcct cgcccttgct caccatggtg
     6661 gcgaccggta atgtcagacc tggtaactcc ttactgtaac ttattgcacc tgatgaacaa
     6721 aagatgtgaa ctgaacgcaa gtaggaggct aatagaggct agtacagtac aaagatcccc
     6781 aaatgatgtg tgccggcctt agtggaggca gacccctggt tatatacctg tgctgcagaa
     6841 agaggggctt acaaatggat ctctctggaa tggcattatc tcatggtggg aacagtgttg
     6901 tcagcagaat cagcggggtg gactctgacc tgagaatgac agtctagcac tgaatggctc
     6961 agacaatact cttgtggcca actatcaccg gaaaataagt gcctgccaaa acaaaccaga
     7021 ccagatgaag ctacatggct gtagaacgtc agtatttaat gtgttgtggc cacttggttc
     7081 accagattta agagctaatt tttttaaata tcacagataa tcaatcgctt ttttttttct
     7141 tgactgaaaa gcttagactg gaaaattctt tgaaaaagtt tgcactacag aaaataaata
     7201 cataacatcc tgcactgaat gcacactatt aatagaaaag agagcattat tgtaggaaat
     7261 gtattgtata tgtacattct gtgcatttca caattttctc gagcaggaaa cagctatgac
     7321 catgattacg ccaagctatc aactttgtat agaaaagttg gtaatacgac tcactatagg
     7381 gcgaattggg taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat
     7441 tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     7501 ttttgttccc tttagtgagg gtta
//
