LOCUS       sequence-178969-        7149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains snx5 specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7149)
  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 7149)
  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;snx5P1Egfp
FEATURES             Location/Qualifiers
     source          1..7149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    10..34
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      315..324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(321..323,324..326,327..1037)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(321..323,324..326,327..1037)
                     /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(366..387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(627..646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     974..995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1038..1103
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1124..1148)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1174..1194)
                     /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(1202..1218)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1234..1368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(1807..1994,1995..2006)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2040..2056)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2093..2111)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2132..2148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2156..2172
                     /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(2180..2186,2187..2204,2205..2210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2225..2246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2363..2380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2534..3122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2614..2633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3293..4084,4085..4153))
                     /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     3916..3935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4154..4258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4421..4442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4791..4802,4803..4965)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5334..5455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5578..6288)
                     /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(5658..5681)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5856..5875)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6125..6143
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6285..6294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     6936..6952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6972..6992
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     6994..7013
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6994..7012
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7021..7128
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7037..7053
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7038..7054
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7088..7104)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7141..7149,1..10))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agggttaatt caagtttgta caaaaaagca ggctcgctat tacatcaaaa ttcttacaag
       61 caacgtttca ttaaatatat tacagttttg taaactgggt gtgtgctatg ttttcgtaaa
      121 tcttttagcg gtgaagtgtc aaaagagtaa aaagtgaaag cgacatttgt tacaagactt
      181 ttattttgac atgatggcag aaaaaatatt cgagccggat gtccacagtg acaggcgaga
      241 gggaggaaag tgaggagcga gcttgctgtt ggaaagttaa aagtgcagtt taatgtacaa
      301 cccagagccg attcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      361 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      421 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      481 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      541 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      601 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      661 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      721 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      781 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
      841 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      901 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
      961 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1021 tggacgagct gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg
     1081 acggtaccgc gggcccggga tccaccggat ctagataact gatacccagc tttcttgtac
     1141 aaagtggggc gcgcctacgt aactagtctc gagcaacttt attatacata gttgataatt
     1201 cactggccgt cgttttacgg taccatcgat gatgatccag acatgataag atacattgat
     1261 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1321 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1381 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1441 aacctctaca aatgtggtat ggctgattat gatcctctag atcagatctg cgaagatacg
     1501 gccacgggtg ctcttgatcc tgtggctgat tttggactgt gctgctcgca gctgctgatg
     1561 aatcacatac ttcctccatt ttcttccact gattgactgt tataatttcc ctaatttcca
     1621 ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga cgtcacttcc aaaggaccaa
     1681 tgaacatgtc tgaccaattt catataatgt gaaaacgatt ttcataggca gaataaataa
     1741 catttaaatt aaactgggca tcagcgcaat tcaattggtt tggtaatagc aagggaaaat
     1801 agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat tgtaaggagt
     1861 aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta ttgattttta
     1921 attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt aatcaagtaa
     1981 aattactcaa gtactttaca cctctggttc ttgaccccct accttcagca agcccagcag
     2041 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     2101 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2161 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2221 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2341 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2941 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3181 aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3601 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3901 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4261 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4321 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4381 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4441 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4501 cccacgctgg ctaagaactc atcagcctcc ccggtccatc tacccacgta ccaatgcacc
     4561 aattggccac aatgacggct actacatggt gccattcctt cctctttata ggaatggaga
     4621 ctacctcctg tccaacaatg ctcttggata cgagtacgcc tacctgttgg acccaggtca
     4681 ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg tgaatatctg ttcagacacc
     4741 catatccact ctgttccaca caggtcagag gtttgtccag gagttcttga cagaggtgta
     4801 aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa attttactca
     4861 attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa agtactccat
     4921 taaaattgta cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa actagagatt
     4981 cttgtttaag cttttaatct caaaaaacat taaatgaaat gcatacaagg ttttatcctg
     5041 ctttagaact gtttgtattt aattatcaaa ctataagaca gacaatctaa tgccagtaca
     5101 cgctactcaa agttgtaaaa cctcagattt aacttcagta gaagctgatt ctcaaaattg
     5161 ttagtgtcaa gcctagctct tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc
     5221 tcacaggcag ccgatgcggg aagaggtgta ttagtcttga tagagaggct gcaaatagca
     5281 ggaaacgtga gcagagactc cctggtgtct gaaacacagg cccatttacg ccttaagata
     5341 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     5401 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     5461 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     5521 caagtaaaac ctctacaaat gtggtatggc tgattatgat ctagagtcgc ggccgcttta
     5581 cttgtacagc tcgtccatgc cgccggtgga gtggtggccc tcggcgcgct cgtactgttc
     5641 cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttgatgttga cgttgtaggc
     5701 gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcgg cgtcgtagtg
     5761 gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg
     5821 gtacatccgc tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc
     5881 ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga
     5941 ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt
     6001 gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta
     6061 ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc
     6121 gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc
     6181 gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat
     6241 gaactccttg atgatggcca tgttgtcatc ctcgcccttg ctcaccatgg tggcgaccgg
     6301 taatgtcaga cctggtaact ccttactgta acttattgca cctgatgaac aaaagatgtg
     6361 aactgaacgc aagtaggagg ctaatagagg ctagtacagt acaaagatcc ccaaatgatg
     6421 tgtgccggcc ttagtggagg cagacccctg gttatatacc tgtgctgcag aaagaggggc
     6481 ttacaaatgg atctctctgg aatggcatta tctcatggtg ggaacagtgt tgtcagcaga
     6541 atcagcgggg tggactctga cctgagaatg acagtctagc actgaatggc tcagacaata
     6601 ctcttgtggc caactatcac cggaaaataa gtgcctgcca aaacaaacca gaccagatga
     6661 agctacatgg ctgtagaacg tcagtattta atgtgttgtg gccacttggt tcaccagatt
     6721 taagagctaa tttttttaaa tatcacagat aatcaatcgc tttttttttt cttgactgaa
     6781 aagcttagac tggaaaattc tttgaaaaag tttgcactac agaaaataaa tacataacat
     6841 cctgcactga atgcacacta ttaatagaaa agagagcatt attgtaggaa atgtattgta
     6901 tatgtacatt ctgtgcattt cacaattttc tcgagcagga aacagctatg accatgatta
     6961 cgccaagcta tcaactttgt atagaaaagt tggtaatacg actcactata gggcgaattg
     7021 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag
     7081 cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc
     7141 cctttagtg
//
