LOCUS       sequence-178857-        7582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains snx5 enhancer sequence, and snx5 specific gene promoter 
            driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  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 7582)
  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-P1Egfp
FEATURES             Location/Qualifiers
     source          1..7582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     641..657
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    677..697
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1297..1321
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1602..1611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1608..1610,1611..1613,1614..2324)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1608..1610,1611..1613,1614..2324)
                     /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(1653..1674)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1914..1933)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2261..2282
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2325..2390
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2411..2435)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2461..2481)
                     /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(2489..2505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2521..2655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3094..3281,3282..3293)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3327..3343)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3327..3343)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3380..3400)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3380..3398)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3419..3435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3432..3454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3443..3459
                     /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(3467..3473,3474..3491,3492..3497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3512..3533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3650..3667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3821..4409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3901..3920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4580..5371,5372..5440))
                     /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     5203..5222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5441..5545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5708..5729)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6078..6089,6090..6252)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6621..6742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6865..7575)
                     /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(6945..6968)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7143..7162)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7412..7430
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7572..7581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 accggtaatg tcagacctgg taactcctta ctgtaactta ttgcacctga tgaacaaaag
       61 atgtgaactg aacgcaagta ggaggctaat agaggctagt acagtacaaa gatccccaaa
      121 tgatgtgtgc cggccttagt ggaggcagac ccctggttat atacctgtgc tgcagaaaga
      181 ggggcttaca aatggatctc tctggaatgg cattatctca tggtgggaac agtgttgtca
      241 gcagaatcag cggggtggac tctgacctga gaatgacagt ctagcactga atggctcaga
      301 caatactctt gtggccaact atcaccggaa aataagtgcc tgccaaaaca aaccagacca
      361 gatgaagcta catggctgta gaacgtcagt atttaatgtg ttgtggccac ttggttcacc
      421 agatttaaga gctaattttt ttaaatatca cagataatca atcgcttttt tttttcttga
      481 ctgaaaagct tagactggaa aattctttga aaaagtttgc actacagaaa ataaatacat
      541 aacatcctgc actgaatgca cactattaat agaaaagaga gcattattgt aggaaatgta
      601 ttgtatatgt acattctgtg catttcacaa ttttctcgag caggaaacag ctatgaccat
      661 gattacgcca agctatcaac tttgtataga aaagttgtat tcctggaata tgaccaggat
      721 gtgagtattc tgctttgtcc aagccaggcc tattcttctc agaccaggaa acagagagac
      781 agtgagaggg actgttggca ttgcttattt cctgcgaagc tgcaatgttg gcagtttaaa
      841 tacaaatggg gagaggttgg aggatgactg cagaagataa agagggaggg catattttac
      901 tcagagaacc acttcctgct tagttcagcg taaaaaaaaa aagattcatg actggactat
      961 tgcttaaaaa aactttaaaa ctaagcctta atctgtaagt cttgtattat tcagataagt
     1021 gtctgttatg ctgataacaa agacaagtga cctttttgga tctgagccag tggtgataaa
     1081 gagcacacac tatacttctg cattctgatt ggttcatatt gtgcgtgact ctgtaatcct
     1141 cttctcattt gggctttagc tcagcgtgag gcggaaaaat gccaaggaaa tgtttggtgg
     1201 tttctttaag aacatggtga agaccgctga tgaggtcctc atctcagggg tcaaggttag
     1261 tgcatcatgc agctctcttt ggggtgtttt taaaatccaa gtttgtacaa aaaagcaggc
     1321 tcgctattac atcaaaattc ttacaagcaa cgtttcatta aatatattac agttttgtaa
     1381 actgggtgtg tgctatgttt tcgtaaatct tttagcggtg aagtgtcaaa agagtaaaaa
     1441 gtgaaagcga catttgttac aagactttta ttttgacatg atggcagaaa aaatattcga
     1501 gccggatgtc cacagtgaca ggcgagaggg aggaaagtga ggagcgagct tgctgttgga
     1561 aagttaaaag tgcagtttaa tgtacaaccc agagccgatt cgccaccatg gtgagcaagg
     1621 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1681 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1741 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1801 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1861 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1921 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1981 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2041 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2101 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2161 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2221 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2281 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc
     2341 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     2401 gataactgat acccagcttt cttgtacaaa gtggggcgcg cctacgtaac tagtctcgag
     2461 caactttatt atacatagtt gataattcac tggccgtcgt tttacggtac catcgatgat
     2521 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2581 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2641 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2701 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     2761 cctctagatc agatctgcga agatacggcc acgggtgctc ttgatcctgt ggctgatttt
     2821 ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc ttccactgat
     2881 tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg taatagatgt
     2941 gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat ataatgtgaa
     3001 aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca gcgcaattca
     3061 attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt
     3121 ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt
     3181 aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa
     3241 taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct ctggttcttg
     3301 accccctacc ttcagcaagc ccagcagatc cactagttct agagcggccg ccaccgcggt
     3361 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     3421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3661 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3721 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4261 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4441 acgctcagtg gaacgaaaac tcacgttaag gcattttggt catgagatta tcaaaaagga
     4501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     5581 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5641 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5701 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5761 gcgaaaaacc gtctatcagg gcgatggccc acgctggcta agaactcatc agcctccccg
     5821 gtccatctac ccacgtacca atgcaccaat tggccacaat gacggctact acatggtgcc
     5881 attccttcct ctttatagga atggagacta cctcctgtcc aacaatgctc ttggatacga
     5941 gtacgcctac ctgttggacc caggtcattg cacaacacca gaaatgccct ctgatctgca
     6001 aaagacgtga atatctgttc agacacccat atccactctg ttccacacag gtcagaggtt
     6061 tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc aagtgaaagt
     6121 acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct agaatcttac
     6181 ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag gaagtaaaag
     6241 taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa aaaacattaa
     6301 atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat tatcaaacta
     6361 taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct cagatttaac
     6421 ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa
     6481 agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag aggtgtatta
     6541 gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct ggtgtctgaa
     6601 acacaggccc atttacgcct taagatacat tgatgagttt ggacaaacca caactagaat
     6661 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6721 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6781 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6841 ttatgatcta gagtcgcggc cgctttactt gtacagctcg tccatgccgc cggtggagtg
     6901 gtggccctcg gcgcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     6961 gtccagcttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
     7021 ggtggtcttg acctcggcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
     7081 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
     7141 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     7201 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     7261 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
     7321 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
     7381 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
     7441 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     7501 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tgtcatcctc
     7561 gcccttgctc accatggtgg cg
//
