LOCUS Exported 7456 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION contains She enhancer sequence and a basal promoter driving expression of eGFP. ACCESSION . VERSION . KEYWORDS pT2-cryR;sheCE1-basEgfp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7456) 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 7456) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7456 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 640..656 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 676..696 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 1143..1167 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1244..1253 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1250..1966 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1250..1966 /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(1295..1316) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1556..1575) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1903..1924 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1967..2032 /label=MCS /note="multiple cloning site" polyA_signal 2156..2277 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(2284..2308) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2334..2354) /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(2362..2378) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 2394..2528 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3200..3216) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3200..3216) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(3253..3273) /label=T3 /note="T3 promoter, forward primer" promoter complement(3253..3271) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3292..3308) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3305..3327) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3316..3332 /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(3340..3370) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3385..3406 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3523..3540) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3694..4282) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3774..3793) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4453..5313) /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 5076..5095 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5314..5418) /gene="bla" /label=AmpR promoter primer_bind complement(5581..5602) /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 6494..6615 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" CDS complement(6738..7448) /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(6818..6841) /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(7016..7035) /label=mCherry-F /note="mCherry, forward primer" primer_bind 7285..7303 /label=mCherry-R /note="mCherry, reverse primer" regulatory 7445..7454 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 ccggtaatgt cagacctggt aactccttac tgtaacttat tgcacctgat gaacaaaaga 61 tgtgaactga acgcaagtag gaggctaata gaggctagta cagtacaaag atccccaaat 121 gatgtgtgcc ggccttagtg gaggcagacc cctggttata tacctgtgct gcagaaagag 181 gggcttacaa atggatctct ctggaatggc attatctcat ggtgggaaca gtgttgtcag 241 cagaatcagc ggggtggact ctgacctgag aatgacagtc tagcactgaa tggctcagac 301 aatactcttg tggccaacta tcaccggaaa ataagtgcct gccaaaacaa accagaccag 361 atgaagctac atggctgtag aacgtcagta tttaatgtgt tgtggccact tggttcacca 421 gatttaagag ctaatttttt taaatatcac agataatcaa tcgctttttt ttttcttgac 481 tgaaaagctt agactggaaa attctttgaa aaagtttgca ctacagaaaa taaatacata 541 acatcctgca ctgaatgcac actattaata gaaaagagag cattattgta ggaaatgtat 601 tgtatatgta cattctgtgc atttcacaat tttctcgagc aggaaacagc tatgaccatg 661 attacgccaa gctatcaact ttgtatagaa aagttgtttg tttaagcaat atcttccttt 721 gctccctctt ctctctgtcg tatctctctt tctctccaca tattctcttc tcattcaaca 781 gctgtattat aagcagacac agtggagctg aaaagcacca aacacaatgt gagccctttc 841 cttgcctctt cctgtctctt ttcctagtca tgaacacact cttcactctg tctttgtggg 901 aaatacactt ttgtgctaca cacacacaca cacacccttg ctcacaaata cgccaagaat 961 acaatgcata cataacacaa tggctctgtc ccattcttcc agtctctggt cagagcagtg 1021 tttgaagaat tatgtcacct gatacatagg ctgtatcctc tcccatttac agacatccct 1081 tccagctaca ctttgaacat aatagcacag aaatcagaat ctcatgaggt cacacacaca 1141 catcaagttt gtacaaaaaa gcaggctaga ctctagaggg tatataatgg atctcgcgtc 1201 tcagcctcac tttgagctcc tccacacgct agcgctaccg gtcgccacca tggtgagcaa 1261 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 1321 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 1381 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 1441 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 1501 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 1561 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 1621 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 1681 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt 1741 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 1801 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 1861 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 1921 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc 1981 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc 2041 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa 2101 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt 2161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 2221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg 2281 cgtacccagc tttcttgtac aaagtggggc gcgcctacgt aactagtctc gagcaacttt 2341 attatacata gttgataatt cactggccgt cgttttacgg taccatcgat gatgatccag 2401 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat 2461 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata 2521 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg 2581 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag 2641 atcagatctg cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt 2701 gctgctcgca gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt 2761 tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga 2821 cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt 2881 ttcataggca gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt 2941 tggtaatagc aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg 3001 taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa 3061 agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact 3121 taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct 3181 accttcagca agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc 3241 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct 3301 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3361 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3421 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3481 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 3541 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 3601 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 3661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 3781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 3841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 3901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 3961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4321 gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac 4381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 4561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 4621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 4681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 4741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 4801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 4861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 4921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat 5461 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 5521 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 5581 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 5641 accgtctatc agggcgatgg cccacgctgg ctaagaactc atcagcctcc ccggtccatc 5701 tacccacgta ccaatgcacc aattggccac aatgacggct actacatggt gccattcctt 5761 cctctttata ggaatggaga ctacctcctg tccaacaatg ctcttggata cgagtacgcc 5821 tacctgttgg acccaggtca ttgcacaaca ccagaaatgc cctctgatct gcaaaagacg 5881 tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag gtttgtccag 5941 gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta 6001 cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct tacttgagta 6061 aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc 6121 aagaaagaaa actagagatt cttgtttaag cttttaatct caaaaaacat taaatgaaat 6181 gcatacaagg ttttatcctg ctttagaact gtttgtattt aattatcaaa ctataagaca 6241 gacaatctaa tgccagtaca cgctactcaa agttgtaaaa cctcagattt aacttcagta 6301 gaagctgatt ctcaaaattg ttagtgtcaa gcctagctct tttggggctg aaaagcaatc 6361 ctgcagtgct gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta ttagtcttga 6421 tagagaggct gcaaatagca ggaaacgtga gcagagactc cctggtgtct gaaacacagg 6481 cccatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga 6541 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 6601 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag 6661 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat 6721 ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgccggtgga gtggtggccc 6781 tcggcgcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc 6841 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc 6901 ttgacctcgg cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc 6961 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc 7021 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg 7081 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag 7141 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg 7201 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc 7261 caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg 7321 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc 7381 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttgtcatc ctcgcccttg 7441 ctcaccatgg tggcga //