LOCUS       sequence-198878-        7645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For miR-223 knock down.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-13- 13"; volume: "22"; issue: "7"; pages: "1810-1823"
COMMENT     Sequence Label: Tol2-UAS-Dendra2-miR-223 sponge
FEATURES             Location/Qualifiers
     source          1..7645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(29..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"
     misc_feature    join(773..784,785..947)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     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"
     misc_feature    join(5170..5357,5358..5369)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     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(join(5543..5549,5550..5567,5568..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(join(6656..7447,7448..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
//
