LOCUS Exported 7645 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For miR-223 knock down. ACCESSION . VERSION . KEYWORDS Tol2-UAS-Dendra2-miR-223 sponge SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7645) 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 7645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7645 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(2..457) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(139..160) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 351..370 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 584..606 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 599..615 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 625..643 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1319..1339 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter 1341..1359 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 1384..1400 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 1385..1401 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1751..1767) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(1804..1822) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" protein_bind 1822..1846 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 1848..1864 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1874..1892 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1936..2628 /codon_start=1 /product="monomeric photoswitchable green-to-red fluorescent protein from Dendronephthya" /label=Dendra2 /note="mammalian codon-optimized" /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY SPLPSQVW" promoter complement(2820..2838) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" protein_bind complement(2867..2891) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" polyA_signal 2896..3030 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2920..2939) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2974..2993 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(5403..5419) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(5456..5474) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5495..5511) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5495..5511) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5508..5530) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5519..5535 /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(5543..5573) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5588..5609 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5726..5743) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5897..6485) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5977..5996) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6656..7516) /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 7279..7298 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7517..7621) /gene="bla" /label=AmpR promoter ORIGIN 1 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca 61 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag 121 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc 181 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc 241 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc 301 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa 361 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc 421 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc 481 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 541 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 601 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacaggt 661 cattgcacaa caccagaaat gccctctgat ctgcaaaaga cgtgaatatc tgttcagaca 721 cccatatcca ctctgttcca cacaggtcag aggtttgtcc aggagttctt gacagaggtg 781 taaaaagtac tcaaaaattt tactcaagtg aaagtacaag tacttaggga aaattttact 841 caattaaaag taaaagtatc tggctagaat cttacttgag taaaagtaaa aaagtactcc 901 attaaaattg tacttgagta ttaaggaagt aaaagtaaaa gcaagaaaga aaactagaga 961 ttcttgttta agcttttaat ctcaaaaaac attaaatgaa atgcatacaa ggttttatcc 1021 tgctttagaa ctgtttgtat ttaattatca aactataaga cagacaatct aatgccagta 1081 cacgctactc aaagttgtaa aacctcagat ttaacttcag tagaagctga ttctcaaaat 1141 tgttagtgtc aagcctagct cttttggggc tgaaaagcaa tcctgcagtg ctgaaaagcc 1201 tctcacaggc agccgatgcg ggaagaggtg tattagtctt gatagagagg ctgcaaatag 1261 caggaaacgt gagcagagac tccctggtgt ctgaaacaca ggccagatgg gccgatatca 1321 actttgtata gaaaagttgg taatacgact cactataggg cgaattgggt accgggcccc 1381 ccctcgaggt cgacggtatc gataagctta ggcctccaag gcggagtact gtcctccggg 1441 ctggcggagt actgtcctcc ggcaaggtcg gagtactgtc ctccgacact agaggtcgga 1501 gtactgtcct ccgacgcaag gcggagtact gtcctccggg ctgcggagta ctgtcctccg 1561 gcaaggtcgg agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg 1621 cggagtactg tcctccgggc tggcggagta ctgtcctccg gcaagggtcg actctagagg 1681 gtatataatg gatcccatcg cgtctcagcc tcactttgag ctcctccaca cgaattcctg 1741 cagcccgggg gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg 1801 ttccctttag tgagggttaa ttcaagtttg tacaaaaaag caggcttgta aaacgacggc 1861 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga 1921 ggtcgactcg ccaccatgaa caccccggga attaacctga tcaaggagga catgcgcgtg 1981 aaggtgcaca tggagggcaa cgtgaacggc cacgccttcg tgatcgaggg cgagggcaag 2041 ggcaagccct acgagggcac ccagaccgcc aacctgaccg tgaaggaggg cgcccccctg 2101 cccttcagct acgacatcct gaccaccgcc gtgcactacg gcaaccgggt gttcaccaag 2161 taccccgagg acatccccga ctacttcaag cagagcttcc ccgagggcta cagctgggag 2221 cgcaccatga ccttcgagga caagggcatc tgcaccatcc gcagcgacat cagcctggag 2281 ggcgactgct tcttccagaa cgtgcgcttc aagggcacca acttcccccc caacggcccc 2341 gtgatgcaga agaagaccct gaagtgggag cccagcaccg agaagctgca cgtgcgcgac 2401 ggcctgctgg tgggcaacat caacatggcc ctgctgctgg agggcggcgg ccactacctg 2461 tgcgacttca agaccaccta caaggccaag aaggtggtgc agctgcccga cgcccacttc 2521 gtggaccacc gcatcgagat cctgggcaac gacagcgact acaacaaggt gaagctgtac 2581 gagcacgccg tggcccgcta cagccccctg cccagccagg tgtggtaagc ggccgctagc 2641 ggggtatttc tgaactgaca ggaaccttgg ggtattttgt aactgacatc tctcgagggg 2701 tatttcgtaa ctgacaaatt cgcgggggta tttcttaact gacagcgcta atggggtatt 2761 ttagaactga cagatcgatc ggggtattta ataactgaca gctagcccag cttttgttcc 2821 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagcaccc agctttcttg 2881 tacaaagttg atgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca 2941 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 3001 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 3061 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg 3121 gccctcgagc atatgagatc tgtcccgatc tttctcttct gtgctgtcaa tttagagtag 3181 tttttgaggt aatttgggtg cattctctgt aaaaacaaca aataaatatt tatattgtat 3241 gcaataaatg taacaattac acatacactc tctgattaca gcattaaagg gttagttcac 3301 ccaaaaatga aaagatgtca ttaatgactc accctcatgt cgttccaagc ccgtaagacc 3361 tccgttcatc ttcagaacac agtttaagat attttagatt tagtccgaga gctttctgtg 3421 cctccattga gaatgtatgt acggtatact gtccatgtcc agaaaggtaa taaaaacatc 3481 aaagtagtcc atgtgacatc agtgggttag ttagaatttt ttgaagcatc gaatacattt 3541 tggtccaaaa ataacaaaac ctacgacttt attcggcatt gtattctctt ccgggtctgt 3601 tgtcaatccg cgttcacgac ttcgcagtga cgctacaatg ctgaataaag tcgtaggttt 3661 tgttattttt ggaccaaaat gtattttcga tgcttcaaat aattctacct aacccactga 3721 tgtcacatgg actactttga tgtttttatt acctttctgg acatggacag tataccgtac 3781 atacattttc agtggaggga cagaaagctc tcggactaaa tctaaaatat cttaaactgt 3841 gttccgaaga tgaacggagg tgttacgggc ttggaacgac atgagggtga gtcattaatg 3901 acatcttttc atttttgggt gaactaaccc tttaataaaa ctgacatttg cagccaattg 3961 cacactttat aatgttaaat acttgttgat tttagaaata cagtagcatt tagtgacaat 4021 caaacattaa ttttatttct gattatattg acaaaaacac aggactgtaa ggctgacatg 4081 acagggtgaa ttgatttcat tacattagtt ttgatggtaa cacttttaca ataaggttcg 4141 ttagttaact acattagtta acatgaacta ataatgaact gcacgtatac atgttaattt 4201 caacatttac taatacttta tatcttgtta acattagtta atgcactgtg aactaacatg 4261 aacaaacaat gaacatctgt atttctatta actaacgtta acaaagatta acagatacag 4321 taacaaatgt attgctcatg gttatttaat gttagttaat acatgaacta atcaacctca 4381 ttgtaaagtg ttaccgtttt gacaatacaa catgtttgga aaaaaatttc tcgcttggtt 4441 ttcttgtgtt ttgcatgaat tcaaacacaa tgagagcacg gaatgataat gattattcaa 4501 agctgcaata ggcgccaaat ttcccgtgga cctaacgtta ccaattacaa atgcatttta 4561 gcattttatt acgtgagcct tatattgtca atcgcatgca ttagtgaaaa taggcctact 4621 acacatataa aaggcgcgcc atcaacgtca aaacatgcac acccaaaaaa aaaaaaaaaa 4681 aaaaaaaaaa aaactatcag taaaacaaaa tacttaatgt acttacctca atatgcttcc 4741 ttaggtttga tggcgaactt ttgaaggccg atatttcttt attaagcggg agacagagga 4801 cacatttcat cttgaatgaa tctttattta caccacttaa agaaaagaat tcgcgaagat 4861 acggccacgg gtgctcttga tcctgtggct gattttggac tgtgctgctc gcagctgctg 4921 atgaatcaca tacttcctcc attttcttcc actgattgac tgttataatt tccctaattt 4981 ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca tgacgtcact tccaaaggac 5041 caatgaacat gtctgaccaa tttcatataa tgtgaaaacg attttcatag gcagaataaa 5101 taacatttaa attaaactgg gcatcagcgc aattcaattg gtttggtaat agcaagggaa 5161 aatagaatga agtgatctcc aaaaaataag tactttttga ctgtaaataa aattgtaagg 5221 agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt 5281 ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac agtaatcaag 5341 taaaattact caagtacttt acacctctgg ttcttgaccc cctaccttca gcaagcccag 5401 cagatccact agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt 5461 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 5641 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 5701 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 5761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 5821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 5881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 5941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 6001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 6061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 6121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 6181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 6241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 6301 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 6361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 6421 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 6481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 6541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 6601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 6661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 6721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 6781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 6841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 6901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 6961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7081 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 7441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 7501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 7561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 7621 gcacatttcc ccgaaaagtg ccacc //