LOCUS       Exported                7923 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  miR-223 expression.
ACCESSION   .
VERSION     .
KEYWORDS    Tol2-lyzC-RFP-miR-223
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7923)
  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 7923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7923
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      1608..1617
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2868..2877
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2874..3581
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKL"
     polyA_signal    3708..3829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3745..3764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3799..3818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4207..4227)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4207..4225)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4246..4262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4246..4262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4259..4281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4270..4286
                     /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(4294..4324)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4339..4360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4477..4494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4648..5236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4728..4747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5407..6267)
                     /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     6030..6049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6268..6372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6535..6556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggttgcatt aagctgcagt tttttcacaa cagttttcca cattgtaaca gtcagtctcc
       61 aaattattcc atcagtcctc aaatttagtc atttgaggcc atgaaaacac ttaaaactac
      121 atgaaggtag tttggcatta ggttttagac ttgttcttgt ttggcatatt gcaacctgtg
      181 taaaaatgat ttgattttta tttttttttt ttttttttca gaaagaaacg gtccaaaaag
      241 agttatgtga acatttatcc atgtgtttaa tgttttttaa attaaaaaaa aacattttta
      301 ttaaaccact tgtgactaaa ccttttttag ttattctatt ttgtgacgag tctgaatggc
      361 caaagtactc ttgaatccaa aatctttata gtaaaatgtt taatcttact tgcagtgttg
      421 ggtataattt gtaacaaagt tgaattccat atctgctgag tgaaaaagta aagtaagaga
      481 ttgctgtaaa tatttaggtt atttatttat taatacaata tagaaagtaa caaaacacaa
      541 tgtaaaaaat caataaagaa agtaaatatt aaaagtaatg ataaactaat gatgtaaaca
      601 atattagcaa caaagtttgt acaattcagg gagggtaaag aatttcacct aatagtcaaa
      661 ctctgtatta aataatataa aatctttgtt cttagtctga agaggagact gtgagagaaa
      721 gcttactggt tgttaaatta gaaaatttgt aacttgcaga ttaaaatgat ttaaataatt
      781 ttataaaggc catgtttgaa acaaatgtaa aaacataata aatgttaaga atttcagata
      841 ttgagatttt acaatttaaa aaaagcaata aagtaatagc agagacctac actcttaaag
      901 taaattaccc aactctgctt actagtaact tttggaatag ttactagtat ccattgaaca
      961 aatactagaa tctaattaaa ttgtgtgagt gtccaatgaa aagattcttg tatttaaaac
     1021 agtaccagta tctatatatg tacaaaattt agtgctagca aaatgcaaat aaatactact
     1081 gcatataatg gattgcattg taaatcgact cgccataata catttatttc ctatttttac
     1141 ttttaacaaa aaataagttc actgagtatt tcatgtcatt aaatcgcatt aaacctcttg
     1201 catgctttat gtacactagt gcagttcgct aaatgaccac cggtgtcact aataacacga
     1261 tcgtttctga attctgcatt aaacgccatt tatacgttgc tcttattagt aaacaatttc
     1321 tccatcatga gctcgaccag tattgcatat ttcatcctct taaatccaga gttgtttctt
     1381 ttaatcactt tctcttcatg ataaaagcag tttttggttt tcagaagttt acaacaaacc
     1441 cagtcagtta ttgcctcagc aaacactgga gcaactgcac acttgtgaaa agcttcttgc
     1501 gaaacacaaa cctttctctg gacttataaa taaaggcatt cagcagctcc accatacacc
     1561 tccactggtc ttgataaagc agatatcagc agtgatacag gtacctcgcc accatggatg
     1621 aggaaatcgc tgccctggtc gttgacaacg gctccggtat gtgcaaagcc ggttttgctg
     1681 gagatgatgc ccctcgtgct gttttcccct ccattgttgg acgacccaga catcaggtac
     1741 atttaataaa ccatcctaat aacatgttaa tcatattgaa ccacattaaa ctctcatttg
     1801 tgcattttac atcttttaaa accgatttat attcgttttt agataatgaa gtttattaaa
     1861 gtgttaattc acagctttat tgcttgtttt gagaatctgc ttgcataagt gatttttaaa
     1921 cttgatctga ttttcgtttg tttcgaagtt cttcagcttt gaaagtcaga tcttcaattc
     1981 acagccatgc catagtttta ggatacagta taagtagacg aattgggatg catcgtagat
     2041 tttatgctgt gtttgttatg gtttttcagt ggtagagatt ttcagagtat tattagggtt
     2101 atagtcagtg tcaggtttat ttatcgtcag cgtgatgggc aaaaaacagc tggcagtacg
     2161 ggctcaggag gaagagggag gagtaaaatt gaatgacctt cctgctgcaa acctgcattt
     2221 aatatccttt tttcctttca tctctcaggt ccatcagcac accccgtctc tctcctcctg
     2281 atctagactc ttctcttaga gtatttgaca gactgtggtt gacactcgat ctaaaggggt
     2341 gtcagtttgt caaatacccc aagagagggg gcacgagtct tcagaattct gcaacacttc
     2401 gtatgaccca tgcgatcctg ttactctctc atgttcagcc tttttgtctt gtgcaaaatg
     2461 tggaggatga caacattcat tagccaactg ctgttactct cgcccagaaa tctgcgccaa
     2521 catggacaca cccagtggag caagccatct gattttcatg gtcagattgt aaagaagcaa
     2581 ttactcttgg acagtttaag cagaaaagcc accaagtcaa gtcactggtc atatgatgaa
     2641 ctcgaagatc taataaaagt taatttaagc gttcattaca ataaattcaa gtgaatctgt
     2701 tttaagctag cctttaacgt ttttaaattc cttcaaacaa attagttcct ctttgcagtt
     2761 cctcatttct cggtgagctg gtgctttaat ccctttactt tttcttcctc tagggagtga
     2821 tggttggcat gggacgaatt aattcctgca gcccgggggt accggtcgcc accatggtgt
     2881 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     2941 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     3001 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     3061 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
     3121 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     3181 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     3241 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     3301 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     3361 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     3421 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     3481 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     3541 ccagatactg cgacctccct agcaaactgg ggcacaaact taagtcgact gcatgcatct
     3601 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3661 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3721 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3781 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3841 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3901 aaccaatagg ccgaaatcgg cggatcctgc ggccgcaagg gaaaatagaa tgaagtgatc
     3961 tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt
     4021 tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta
     4081 aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac
     4141 tttacacctc tggttcttga ccccctacct tcagcaagcc cagcagatcc actagatctt
     4201 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4261 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4321 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4441 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6001 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6361 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6421 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6481 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6541 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6601 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6661 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6721 aagccggcca caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag
     6781 actacctcct gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc
     6841 attgcacaac accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac
     6901 ccatatccac tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt
     6961 aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc
     7021 aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca
     7081 ttaaaattgt acttgagtat taaggaagta aaaggctagc aagatctgct cgagatacat
     7141 ttgattagaa gagtttgact tgggttgatt atagcaaata gtggaacaca aacagatttt
     7201 gaaatattct tgcaaattta tttattaaat ccattcaaat attgtcattt tttgtactgt
     7261 gtaatttata ttgatttgct catacaatat ttacatttcg cttaaacaat acactattat
     7321 ttatatttaa aaatagctat atataaatca aatgtatagc tcttttttac acaattaatc
     7381 aaagctaaaa tatttgtcac acaaatacag taagccactt cacaatgata tcttcaaaat
     7441 ctgtgcttga ctgaatcatt tcatttattg ttttgtaatt cacattatac cttacgttct
     7501 ccaggcacag tttgaattgt tgataaatga agataataca tctgatttaa ttagtctcct
     7561 ttgtttataa taactttaat ctaatttgat agtaataatt ccagcaatac atccattaca
     7621 ataaaaatcg cttttttatt ttgcacatta gtttgccggt atttgtttaa agtttcacca
     7681 tacttttaaa tcatgtgcta tttgaaaagg gctgttatga ctggaattaa atgttttttt
     7741 ttatatcagt gttttagatt gaagtacaat atttataggt aaaattcacc agcatatgct
     7801 taaagatagc catgctccag gtccttgtgt ccttctaagg tctgtttgtg gtttaagctg
     7861 tttgttgaca cacgcttgtc ttctccattt ttctgtgaag ttattgtcag tttattgcac
     7921 act
//
