LOCUS Exported 9298 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For Gal4/UAS expression system. ACCESSION . VERSION . KEYWORDS Tol2-lyzC-Gal4-Vp16-Crys-CFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9298) AUTHORS Zhou W, Pal AS, Hsu AY, Gurol T, Zhu X, Wirbisky-Hershberger SE, Freeman JL, Kasinski AL, Deng Q TITLE MicroRNA-223 Suppresses the Canonical NF-kappaB Pathway in Basal Keratinocytes to Dampen Neutrophilic Inflammation. JOURNAL Cell Rep. 2018 Feb 13;22(7):1810-1823. doi: 10.1016/j.celrep.2018.01.058. PUBMED 29444433 REFERENCE 2 (bases 1 to 9298) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9298 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1710..1734 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 1735..1752 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1736..1752 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1762..1781 /label=T7 /note="T7 promoter, forward primer" promoter 1762..1780 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1801..2238 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /label=GAL4 DNA binding domain /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTV" promoter complement(2470..2487) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2492..2626 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(2723..2741) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" protein_bind complement(2770..2794) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind 3445..3478 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." regulatory 3631..3640 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3637..4356 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3682..3703) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3943..3962) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4290..4311 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(4385..4405) /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(4413..4429) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 4445..4579 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5251..5267) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5251..5267) /label=pBluescriptSK /note="For pBluescript vector" promoter complement(5304..5322) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5343..5359) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5356..5378) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5367..5383 /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(5391..5421) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5436..5457 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5574..5591) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5745..6333) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5825..5844) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6504..7364) /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 7127..7146 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7365..7469) /gene="bla" /label=AmpR promoter primer_bind complement(7632..7653) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8549..8565 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 8585..8605 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" ORIGIN 1 caggtccttg tgtccttcta aggtctgttt gtggtttaag ctgtttgttg acacacgctt 61 gtcttctcca tttttctgtg aagttattgt cagtttattg cacactgggt tgcattaagc 121 tgcagttttt tcacaacagt tttccacatt gtaacagtca gtctccaaat tattccatca 181 gtcctcaaat ttagtcattt gaggccatga aaacacttaa aactacatga aggtagtttg 241 gcattaggtt ttagacttgt tcttgtttgg catattgcaa cctgtgtaaa aatgatttga 301 tttttatttt tttttttttt ttttcagaaa gaaacggtcc aaaaagagtt atgtgaacat 361 ttatccatgt gtttaatgtt ttttaaatta aaaaaaaaca tttttattaa accacttgtg 421 actaaacctt ttttagttat tctattttgt gacgagtctg aatggccaaa gtactcttga 481 atccaaaatc tttatagtaa aatgtttaat cttacttgca gtgttgggta taatttgtaa 541 caaagttgaa ttccatatct gctgagtgaa aaagtaaagt aagagattgc tgtaaatatt 601 taggttattt atttattaat acaatataga aagtaacaaa acacaatgta aaaaatcaat 661 aaagaaagta aatattaaaa gtaatgataa actaatgatg taaacaatat tagcaacaaa 721 gtttgtacaa ttcagggagg gtaaagaatt tcacctaata gtcaaactct gtattaaata 781 atataaaatc tttgttctta gtctgaagag gagactgtga gagaaagctt actggttgtt 841 aaattagaaa atttgtaact tgcagattaa aatgatttaa ataattttat aaaggccatg 901 tttgaaacaa atgtaaaaac ataataaatg ttaagaattt cagatattga gattttacaa 961 tttaaaaaaa gcaataaagt aatagcagag acctacactc ttaaagtaaa ttacccaact 1021 ctgcttacta gtaacttttg gaatagttac tagtatccat tgaacaaata ctagaatcta 1081 attaaattgt gtgagtgtcc aatgaaaaga ttcttgtatt taaaacagta ccagtatcta 1141 tatatgtaca aaatttagtg ctagcaaaat gcaaataaat actactgcat ataatggatt 1201 gcattgtaaa tcgactcgcc ataatacatt tatttcctat ttttactttt aacaaaaaat 1261 aagttcactg agtatttcat gtcattaaat cgcattaaac ctcttgcatg ctttatgtac 1321 actagtgcag ttcgctaaat gaccaccggt gtcactaata acacgatcgt ttctgaattc 1381 tgcattaaac gccatttata cgttgctctt attagtaaac aatttctcca tcatgagctc 1441 gaccagtatt gcatatttca tcctcttaaa tccagagttg tttcttttaa tcactttctc 1501 ttcatgataa aagcagtttt tggttttcag aagtttacaa caaacccagt cagttattgc 1561 ctcagcaaac actggagcaa ctgcacactt gtgaaaagct tcttgcgaaa cacaaacctt 1621 tctctggact tataaataaa ggcattcagc agctccacca tacacctcca ctggtcttga 1681 taaagggatc ctttaagggc gaattcgacc caagtttgta caaaaaagca ggcttgtaaa 1741 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgccacc 1801 atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag 1861 tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac 1921 tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg 1981 ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt 2041 ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat 2101 aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta 2161 acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt 2221 caaagacagt tgactgtatc ttcgaggtcg accccgggaa ttcagatctc tcgagccgcc 2281 cccccgaccg atgtcagcct gggggacgag ctccacttag acggcgagga cgtggcgatg 2341 gcgcatgccg acgcgctaga cgatttcgat ctggacatgt tgggggacgg ggattccccg 2401 ggtccgggat ttacccccca cgactccgcc ccctacggcg ctctggatat gtagctcgag 2461 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga 2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 2641 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc 2701 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat 2761 ggtcatagca cccagctttc ttgtacaaag tggaccgtcg agcaattggt gctatcatca 2821 tgacaaagag agaagaaatc agttattaga aatgagttat taaaactatt atgattagaa 2881 atatgtttct ttctcttaaa caggatttaa ggggaaaata tacagggagc gaaaaatttt 2941 gacttcatct gtatatacat ataataaata atacatcaat aaattccatc taaatacgct 3001 accatatata tatatatatg tatatacaca cacacacaca caccatctaa tatatgtcat 3061 gctacattat taacttcagt atgaaatcta ctctgctctg gagtgtatca gcgagcgact 3121 cctctgctcg gattatcatc acgtgacctc tgacctgaac acagacctct cttagccata 3181 ggtatacagc ctatagcata gagctcgggc tgagagggaa agtgccgccg tcagggcttt 3241 ccattgacag taaaatgctt acggccttct gggcgatgcg atgaccctgc gcttcggcac 3301 ggctgaactc tgatgacggc gtgtttctat tgtccgccgc gcgccgctgt gctgcccttg 3361 ccagcgtata aaagctgcgg cagtggtgag gaaggccaga gcgagccagc cttcccagca 3421 cagctgtagt ttaaacgcgt tgccataact tcgtatagga tactttatac gaagttatct 3481 gcatgctagg ttgccgggaa cggtgcattg gaacgcgcat tccccgtgtt aattaacagg 3541 taagtgtctt cctcctgttt ccttcccctg ctattctgct caaccttcct atcagaaact 3601 gcagtatctg tatttttgct aggggatccc gccaccatgg tgagcaaggg cgaggagctg 3661 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 3721 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 3781 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc 3841 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 3901 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 3961 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 4021 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc 4081 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc 4141 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 4201 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 4261 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 4321 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccaaacccg ctgatcagcc 4381 tcgacaactt tattatacat agttgataat tcactggccg tcgttttacg gtaccatcga 4441 tgatgatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 4501 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 4561 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg 4621 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta 4681 tgatcctcta gatcagatct gcgaagatac ggccacgggt gctcttgatc ctgtggctga 4741 ttttggactg tgctgctcgc agctgctgat gaatcacata cttcctccat tttcttccac 4801 tgattgactg ttataatttc cctaatttcc aggtcaaggt gctgtgcatt gtggtaatag 4861 atgtgacatg acgtcacttc caaaggacca atgaacatgt ctgaccaatt tcatataatg 4921 tgaaaacgat tttcataggc agaataaata acatttaaat taaactgggc atcagcgcaa 4981 ttcaattggt ttggtaatag caagggaaaa tagaatgaag tgatctccaa aaaataagta 5041 ctttttgact gtaaataaaa ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt 5101 aattaagtaa aagtaaaagt attgattttt aattgtactc aagtaaagta aaaatcccca 5161 aaaataatac ttaagtacag taatcaagta aaattactca agtactttac acctctggtt 5221 cttgaccccc taccttcagc aagcccagca gatccactag ttctagagcg gccgccaccg 5281 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca 5341 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5401 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5461 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6361 tctgacgctc agtggaacga aaactcacgt taaggcattt tggtcatgag attatcaaaa 6421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg 7501 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 7561 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 7621 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 7681 aagggcgaaa aaccgtctat cagggcgatg gcccacgctg gctaagaact catcagcctc 7741 cccggtccat ctacccacgt accaatgcac caattggcca caatgacggc tactacatgg 7801 tgccattcct tcctctttat aggaatggag actacctcct gtccaacaat gctcttggat 7861 acgagtacgc ctacctgttg gacccaggtc attgcacaac accagaaatg ccctctgatc 7921 tgcaaaagac gtgaatatct gttcagacac ccatatccac tctgttccac acaggtcaga 7981 ggtttgtcca ggagttcttg acagaggtgt aaaaagtact caaaaatttt actcaagtga 8041 aagtacaagt acttagggaa aattttactc aattaaaagt aaaagtatct ggctagaatc 8101 ttacttgagt aaaagtaaaa aagtactcca ttaaaattgt acttgagtat taaggaagta 8161 aaagtaaaag caagaaagaa aactagagat tcttgtttaa gcttttaatc tcaaaaaaca 8221 ttaaatgaaa tgcatacaag gttttatcct gctttagaac tgtttgtatt taattatcaa 8281 actataagac agacaatcta atgccagtac acgctactca aagttgtaaa acctcagatt 8341 taacttcagt agaagctgat tctcaaaatt gttagtgtca agcctagctc ttttggggct 8401 gaaaagcaat cctgcagtgc tgaaaagcct ctcacaggca gccgatgcgg gaagaggtgt 8461 attagtcttg atagagaggc tgcaaatagc aggaaacgtg agcagagact ccctggtgtc 8521 tgaaacacag gccagatggg ccctcgagca ggaaacagct atgaccatga ttacgccaag 8581 ctatcaactt tgtatagaaa agttggctcc gaattcgccc ttaaaactcg agagtttgac 8641 ttgggttgat tatagcaaat agtggaacac aaacagattt tgaaatattc ttgcaaattt 8701 atttattaaa tccattcaaa tattgtcatt ttttgtactg tgtaatttat attgatttgc 8761 tcatacaata tttacatttc gcttaaacaa tacactatta tttatattta aaaatagcta 8821 tatataaatc aaatgtatag ctctttttta cacaattaat caaagctaaa atatttgtca 8881 cacaaataca gtaagccact tcacaatgat atcttcaaaa tctgtgcttg actgaatcat 8941 ttcatttatt gttttgtaat tcacattata ccttacgttc tccaggcaca gtttgaattg 9001 ttgataaatg aagataatac atctgattta attagtctcc tttgtttata ataactttaa 9061 tctaatttga tagtaataat tccagcaata catccattac aataaaaatc gcttttttat 9121 tttgcacatt agtttgccgg tatttgttta aagtttcacc atacttttaa atcatgtgct 9181 atttgaaaag ggctgttatg actggaatta aatgtttttt tttatatcag tgttttagat 9241 tgaagtacaa tatttatagg taaaattcac cagcatatgc ttaaagatag ccatgctc //