LOCUS       sequence-178954-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains lmo2 specific gene promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  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 7534)
  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;lmo2P1Egfp
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..29
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      695..704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(701..703,704..706,707..1417)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(701..703,704..706,707..1417)
                     /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(746..767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1007..1026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1354..1375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1418..1483
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1504..1528)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1554..1574)
                     /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(1582..1598)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1614..1748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(2187..2374,2375..2386)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2420..2436)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2473..2491)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2512..2528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2525..2547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2536..2552
                     /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(2560..2566,2567..2584,2585..2590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2605..2626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2743..2760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2914..3502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2994..3013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3673..4464,4465..4533))
                     /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     4296..4315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4534..4638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4801..4822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5171..5182,5183..5345)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5714..5835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5958..6668)
                     /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(6038..6061)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6236..6255)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6505..6523
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6665..6674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     7316..7332
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7352..7372
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     7374..7393
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7374..7392
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7401..7508
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7417..7433
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7418..7434
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7468..7484)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(7521..7534,1..5))
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 taattcaagt ttgtacaaaa aagcaggctc gctactgctc cagcaagagt tctgaataca
       61 taaaaaagta aaatgattgt ttaaccctaa aagcaattat aaaacaatag gcctatttgc
      121 caacaagaca aaatgtggag agttcttttt aaggggttca tccaaaaaaa aatctgatta
      181 tgtcatttac ttttattgtt gcaaactgtt gaacacaaac aaaaagttgc aatgaatcct
      241 gaaaaccagt attggcttcc aaaatagcct attgttacag ttttctaaca ttcctcacaa
      301 taactttgtc aagagaacaa aacgaaatgt ttgccagcca agttgtttaa cctcctcgtt
      361 tggatgcagc caggtatttg atttctgcct gactgactcc gtgtttgagc tcgtgacccc
      421 ttaagtcacg agtcgtcggt gtcggctgta taattgactg gtgattcgcg catgtaggag
      481 cgctatttcc tctcattgtc gagccgtcgt gagcaggaag tcggaaaaga gcagaaactg
      541 agggggcagg gccatttaaa aatacctgct gccctacaat aacgggacac agagacagca
      601 gccctgcggg acactggcgt ggcgacttgc agcgcaaaat gcaccacaaa caagacggag
      661 cctaaacgcc acatttatca cgcgccggag aatcgccacc atggtgagca agggcgagga
      721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1381 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
     1441 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     1501 gatacccagc tttcttgtac aaagtggggc gcgcctacgt aactagtctc gagcaacttt
     1561 attatacata gttgataatt cactggccgt cgttttacgg taccatcgat gatgatccag
     1621 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1681 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1741 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1801 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag
     1861 atcagatctg cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt
     1921 gctgctcgca gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt
     1981 tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga
     2041 cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt
     2101 ttcataggca gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt
     2161 tggtaatagc aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg
     2221 taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa
     2281 agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact
     2341 taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct
     2401 accttcagca agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc
     2461 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2521 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2581 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2641 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2701 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3541 gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3781 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3961 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4261 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4501 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4561 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4621 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     4681 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4741 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4801 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4861 accgtctatc agggcgatgg cccacgctgg ctaagaactc atcagcctcc ccggtccatc
     4921 tacccacgta ccaatgcacc aattggccac aatgacggct actacatggt gccattcctt
     4981 cctctttata ggaatggaga ctacctcctg tccaacaatg ctcttggata cgagtacgcc
     5041 tacctgttgg acccaggtca ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg
     5101 tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag gtttgtccag
     5161 gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta
     5221 cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct tacttgagta
     5281 aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc
     5341 aagaaagaaa actagagatt cttgtttaag cttttaatct caaaaaacat taaatgaaat
     5401 gcatacaagg ttttatcctg ctttagaact gtttgtattt aattatcaaa ctataagaca
     5461 gacaatctaa tgccagtaca cgctactcaa agttgtaaaa cctcagattt aacttcagta
     5521 gaagctgatt ctcaaaattg ttagtgtcaa gcctagctct tttggggctg aaaagcaatc
     5581 ctgcagtgct gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta ttagtcttga
     5641 tagagaggct gcaaatagca ggaaacgtga gcagagactc cctggtgtct gaaacacagg
     5701 cccatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     5761 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     5821 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     5881 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     5941 ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgccggtgga gtggtggccc
     6001 tcggcgcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc
     6061 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc
     6121 ttgacctcgg cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc
     6181 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc
     6241 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg
     6301 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag
     6361 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg
     6421 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc
     6481 caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg
     6541 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc
     6601 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttgtcatc ctcgcccttg
     6661 ctcaccatgg tggcgaccgg taatgtcaga cctggtaact ccttactgta acttattgca
     6721 cctgatgaac aaaagatgtg aactgaacgc aagtaggagg ctaatagagg ctagtacagt
     6781 acaaagatcc ccaaatgatg tgtgccggcc ttagtggagg cagacccctg gttatatacc
     6841 tgtgctgcag aaagaggggc ttacaaatgg atctctctgg aatggcatta tctcatggtg
     6901 ggaacagtgt tgtcagcaga atcagcgggg tggactctga cctgagaatg acagtctagc
     6961 actgaatggc tcagacaata ctcttgtggc caactatcac cggaaaataa gtgcctgcca
     7021 aaacaaacca gaccagatga agctacatgg ctgtagaacg tcagtattta atgtgttgtg
     7081 gccacttggt tcaccagatt taagagctaa tttttttaaa tatcacagat aatcaatcgc
     7141 tttttttttt cttgactgaa aagcttagac tggaaaattc tttgaaaaag tttgcactac
     7201 agaaaataaa tacataacat cctgcactga atgcacacta ttaatagaaa agagagcatt
     7261 attgtaggaa atgtattgta tatgtacatt ctgtgcattt cacaattttc tcgagcagga
     7321 aacagctatg accatgatta cgccaagcta tcaactttgt atagaaaagt tggtaatacg
     7381 actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc
     7441 ttgatatcga attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg
     7501 tggagctcca gcttttgttc cctttagtga gggt
//
