LOCUS       sequence-178706-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains snx5 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 7609)
  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 7609)
  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;snx5CE1-basEgfp
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     646..662
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    682..702
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1302..1326
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1403..1412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1409..1411,1412..1414,1415..2125)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1409..1411,1412..1414,1415..2125)
                     /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(1454..1475)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1715..1734)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2062..2083
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2126..2191
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2315..2436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2443..2467)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2493..2513)
                     /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(2521..2537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2553..2687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3126..3313,3314..3325)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3359..3375)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3359..3375)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3412..3432)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3412..3430)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3451..3467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3464..3486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3475..3491
                     /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(3499..3505,3506..3523,3524..3529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3544..3565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3682..3699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3853..4441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3933..3952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4612..5403,5404..5472))
                     /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     5235..5254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5473..5577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5740..5761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6110..6121,6122..6284)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6653..6774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6897..7607)
                     /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(6977..7000)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7175..7194)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7444..7462
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      join(7604..7609,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tggcgaccgg taatgtcaga cctggtaact ccttactgta acttattgca cctgatgaac
       61 aaaagatgtg aactgaacgc aagtaggagg ctaatagagg ctagtacagt acaaagatcc
      121 ccaaatgatg tgtgccggcc ttagtggagg cagacccctg gttatatacc tgtgctgcag
      181 aaagaggggc ttacaaatgg atctctctgg aatggcatta tctcatggtg ggaacagtgt
      241 tgtcagcaga atcagcgggg tggactctga cctgagaatg acagtctagc actgaatggc
      301 tcagacaata ctcttgtggc caactatcac cggaaaataa gtgcctgcca aaacaaacca
      361 gaccagatga agctacatgg ctgtagaacg tcagtattta atgtgttgtg gccacttggt
      421 tcaccagatt taagagctaa tttttttaaa tatcacagat aatcaatcgc tttttttttt
      481 cttgactgaa aagcttagac tggaaaattc tttgaaaaag tttgcactac agaaaataaa
      541 tacataacat cctgcactga atgcacacta ttaatagaaa agagagcatt attgtaggaa
      601 atgtattgta tatgtacatt ctgtgcattt cacaattttc tcgagcagga aacagctatg
      661 accatgatta cgccaagcta tcaactttgt atagaaaagt tgtattcctg gaatatgacc
      721 aggatgtgag tattctgctt tgtccaagcc aggcctattc ttctcagacc aggaaacaga
      781 gagacagtga gagggactgt tggcattgct tatttcctgc gaagctgcaa tgttggcagt
      841 ttaaatacaa atggggagag gttggaggat gactgcagaa gataaagagg gagggcatat
      901 tttactcaga gaaccacttc ctgcttagtt cagcgtaaaa aaaaaaagat tcatgactgg
      961 actattgctt aaaaaaactt taaaactaag ccttaatctg taagtcttgt attattcaga
     1021 taagtgtctg ttatgctgat aacaaagaca agtgaccttt ttggatctga gccagtggtg
     1081 ataaagagca cacactatac ttctgcattc tgattggttc atattgtgcg tgactctgta
     1141 atcctcttct catttgggct ttagctcagc gtgaggcgga aaaatgccaa ggaaatgttt
     1201 ggtggtttct ttaagaacat ggtgaagacc gctgatgagg tcctcatctc aggggtcaag
     1261 gttagtgcat catgcagctc tctttggggt gtttttaaaa tccaagtttg tacaaaaaag
     1321 caggctagac tctagagggt atataatgga tctcgcgtct cagcctcact ttgagctcct
     1381 ccacacgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1441 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1501 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1561 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1621 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1681 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1741 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1801 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1861 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1921 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1981 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2041 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2101 tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgaattc
     2161 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     2221 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2281 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2341 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2401 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtacccagct ttcttgtaca
     2461 aagtggggcg cgcctacgta actagtctcg agcaacttta ttatacatag ttgataattc
     2521 actggccgtc gttttacggt accatcgatg atgatccaga catgataaga tacattgatg
     2581 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2641 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2701 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2761 acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctgc gaagatacgg
     2821 ccacgggtgc tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga
     2881 atcacatact tcctccattt tcttccactg attgactgtt ataatttccc taatttccag
     2941 gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat
     3001 gaacatgtct gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac
     3061 atttaaatta aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata
     3121 gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta
     3181 aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa
     3241 ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa
     3301 attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga
     3361 tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     3421 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4501 aggcattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5581 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5641 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5701 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5761 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5821 ccacgctggc taagaactca tcagcctccc cggtccatct acccacgtac caatgcacca
     5881 attggccaca atgacggcta ctacatggtg ccattccttc ctctttatag gaatggagac
     5941 tacctcctgt ccaacaatgc tcttggatac gagtacgcct acctgttgga cccaggtcat
     6001 tgcacaacac cagaaatgcc ctctgatctg caaaagacgt gaatatctgt tcagacaccc
     6061 atatccactc tgttccacac aggtcagagg tttgtccagg agttcttgac agaggtgtaa
     6121 aaagtactca aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa
     6181 ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa gtactccatt
     6241 aaaattgtac ttgagtatta aggaagtaaa agtaaaagca agaaagaaaa ctagagattc
     6301 ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg catacaaggt tttatcctgc
     6361 tttagaactg tttgtattta attatcaaac tataagacag acaatctaat gccagtacac
     6421 gctactcaaa gttgtaaaac ctcagattta acttcagtag aagctgattc tcaaaattgt
     6481 tagtgtcaag cctagctctt ttggggctga aaagcaatcc tgcagtgctg aaaagcctct
     6541 cacaggcagc cgatgcggga agaggtgtat tagtcttgat agagaggctg caaatagcag
     6601 gaaacgtgag cagagactcc ctggtgtctg aaacacaggc ccatttacgc cttaagatac
     6661 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     6721 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     6781 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     6841 aagtaaaacc tctacaaatg tggtatggct gattatgatc tagagtcgcg gccgctttac
     6901 ttgtacagct cgtccatgcc gccggtggag tggtggccct cggcgcgctc gtactgttcc
     6961 acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tgatgttgac gttgtaggcg
     7021 ccgggcagct gcacgggctt cttggccttg taggtggtct tgacctcggc gtcgtagtgg
     7081 ccgccgtcct tcagcttcag cctctgcttg atctcgccct tcagggcgcc gtcctcgggg
     7141 tacatccgct cggaggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg
     7201 gaggggaagt tggtgccgcg cagcttcacc ttgtagatga actcgccgtc ctgcagggag
     7261 gagtcctggg tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg
     7321 aagccctcgg ggaaggacag cttcaagtag tcggggatgt cggcggggtg cttcacgtag
     7381 gccttggagc cgtacatgaa ctgaggggac aggatgtccc aggcgaaggg cagggggccg
     7441 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     7501 ccctcgatct cgaactcgtg gccgttcacg gagccctcca tgtgcacctt gaagcgcatg
     7561 aactccttga tgatggccat gttgtcatcc tcgcccttgc tcaccatgg
//
