LOCUS       sequence-198866-        7516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Gal4/UAS expression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7516)
  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 7516)
  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
FEATURES             Location/Qualifiers
     source          1..7516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(14..469)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(151..172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     363..382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     596..618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     611..627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(785..796,797..959)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    1331..1351
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1353..1371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1396..1412
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1397..1413
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1763..1779)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1816..1834)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1834..1858
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1860..1876
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1886..1904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1948..2640
                     /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"
     primer_bind     complement(2657..2673)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2704..2722)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2751..2775)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2780..2914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2804..2823)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2858..2877
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5053..5240,5241..5252)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5286..5302)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5339..5357)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5378..5394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5378..5394)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5391..5413)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5402..5418
                     /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(5426..5432,5433..5450,5451..5456))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5471..5492
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5609..5626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5780..6368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5860..5879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6539..7330,7331..7399))
                     /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     7162..7181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7400..7504)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
       61 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      121 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      181 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      241 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      301 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      361 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      421 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
      481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      601 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
      661 ttgggtacag gtcattgcac aacaccagaa atgccctctg atctgcaaaa gacgtgaata
      721 tctgttcaga cacccatatc cactctgttc cacacaggtc agaggtttgt ccaggagttc
      781 ttgacagagg tgtaaaaagt actcaaaaat tttactcaag tgaaagtaca agtacttagg
      841 gaaaatttta ctcaattaaa agtaaaagta tctggctaga atcttacttg agtaaaagta
      901 aaaaagtact ccattaaaat tgtacttgag tattaaggaa gtaaaagtaa aagcaagaaa
      961 gaaaactaga gattcttgtt taagctttta atctcaaaaa acattaaatg aaatgcatac
     1021 aaggttttat cctgctttag aactgtttgt atttaattat caaactataa gacagacaat
     1081 ctaatgccag tacacgctac tcaaagttgt aaaacctcag atttaacttc agtagaagct
     1141 gattctcaaa attgttagtg tcaagcctag ctcttttggg gctgaaaagc aatcctgcag
     1201 tgctgaaaag cctctcacag gcagccgatg cgggaagagg tgtattagtc ttgatagaga
     1261 ggctgcaaat agcaggaaac gtgagcagag actccctggt gtctgaaaca caggccagat
     1321 gggccgatat caactttgta tagaaaagtt ggtaatacga ctcactatag ggcgaattgg
     1381 gtaccgggcc ccccctcgag gtcgacggta tcgataagct taggcctcca aggcggagta
     1441 ctgtcctccg ggctggcgga gtactgtcct ccggcaaggt cggagtactg tcctccgaca
     1501 ctagaggtcg gagtactgtc ctccgacgca aggcggagta ctgtcctccg ggctgcggag
     1561 tactgtcctc cggcaaggtc ggagtactgt cctccgacac tagaggtcgg agtactgtcc
     1621 tccgacgcaa ggcggagtac tgtcctccgg gctggcggag tactgtcctc cggcaagggt
     1681 cgactctaga gggtatataa tggatcccat cgcgtctcag cctcactttg agctcctcca
     1741 cacgaattcc tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag
     1801 ctccagcttt tgttcccttt agtgagggtt aattcaagtt tgtacaaaaa agcaggcttg
     1861 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     1921 gccccccctc gaggtcgact cgccaccatg aacaccccgg gaattaacct gatcaaggag
     1981 gacatgcgcg tgaaggtgca catggagggc aacgtgaacg gccacgcctt cgtgatcgag
     2041 ggcgagggca agggcaagcc ctacgagggc acccagaccg ccaacctgac cgtgaaggag
     2101 ggcgcccccc tgcccttcag ctacgacatc ctgaccaccg ccgtgcacta cggcaaccgg
     2161 gtgttcacca agtaccccga ggacatcccc gactacttca agcagagctt ccccgagggc
     2221 tacagctggg agcgcaccat gaccttcgag gacaagggca tctgcaccat ccgcagcgac
     2281 atcagcctgg agggcgactg cttcttccag aacgtgcgct tcaagggcac caacttcccc
     2341 cccaacggcc ccgtgatgca gaagaagacc ctgaagtggg agcccagcac cgagaagctg
     2401 cacgtgcgcg acggcctgct ggtgggcaac atcaacatgg ccctgctgct ggagggcggc
     2461 ggccactacc tgtgcgactt caagaccacc tacaaggcca agaaggtggt gcagctgccc
     2521 gacgcccact tcgtggacca ccgcatcgag atcctgggca acgacagcga ctacaacaag
     2581 gtgaagctgt acgagcacgc cgtggcccgc tacagccccc tgcccagcca ggtgtggtaa
     2641 gagctgtaca agtaaggatc cactagttct agagcggccg cggtggagct ccagcttttg
     2701 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc acccagcttt
     2761 cttgtacaaa gttgatgtag atccagacat gataagatac attgatgagt ttggacaaac
     2821 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2881 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2941 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     3001 ttgggccctc gagcatatga gatctgtccc gatctttctc ttctgtgctg tcaatttaga
     3061 gtagtttttg aggtaatttg ggtgcattct ctgtaaaaac aacaaataaa tatttatatt
     3121 gtatgcaata aatgtaacaa ttacacatac actctctgat tacagcatta aagggttagt
     3181 tcacccaaaa atgaaaagat gtcattaatg actcaccctc atgtcgttcc aagcccgtaa
     3241 cacctccgtt catcttcgga acacagttta agatatttta gatttagtcc gagagctttc
     3301 tgtccctcca ctgaaaatgt atgtacggta tactgtccat gtccagaaag gtaataaaaa
     3361 catcaaagta gtccatgtga catcagtggg ttagttagaa ttttttgaag catcgaatac
     3421 attttggtcc aaaaataaca aaacctacga ctttattcgg cattgtattc tcttccgggt
     3481 ctgttgtcaa tccgcgttca cgacttcgca gtgacgctac aatgctgaat aaagtcgtag
     3541 gttttgttat ttttggacca aaatgtattt tcgatgcttc aaataattct acctaaccca
     3601 ctgatgtcac atggactact ttgatgtttt tattaccttt ctggacatgg acagtatacc
     3661 gtacatacat tctcaatgga ggcacagaaa gctctcggac taaatctaaa atatcttaaa
     3721 ctgtgttctg aagatgaacg gaggtcttac gggcttggaa cgacatgagg gcgagtcatt
     3781 aatgacatta ttttcatttt tgggtgaact aaccctttaa taaaactgac atttgcagcc
     3841 aattgcacac tttataatgt taaatacttg ttgattttag aaatacagta gcatttagtg
     3901 acaatcaaac attaatttta tttctgatta tattgacaaa aacacaggac tgtaaggctg
     3961 acatgacagg gtgaattgat ttcattacat tagttttgat ggtaacactt ttacaataag
     4021 gttcgttagt taactacatt agttaacatg aactaataat gaactgcacg tatacatgtt
     4081 aatttcaaca tttactaata ctttatatct tgttaacatt agttaatgca ctgtgaacta
     4141 acatgaacaa acaatgaaca tctgtatttc tattaactaa cgttaacaaa gattaacaga
     4201 tacagtaaca aatgtattgc tcatggttat ttaatgttag ttaatacatg aactaatcaa
     4261 cctcattgta aagtgttacc gttttgacaa tacaacatgt ttggaaaaaa atttctcgct
     4321 tggttttctt gtgttttgca tgaattcaaa cacaatgaga gcacggaatg ataatgatta
     4381 ttcaaagctg caataggcgc caaatttccc gtggacctaa cgttaccaat tacaaatgca
     4441 ttttagcatt ttattacgtg agccttatat tgtcaatcgc atgcattagt gaaaataggc
     4501 ctactacaca tataaaaggc gcgccatcaa cgtcaaaaca tgcacaccca aaaaaaaaaa
     4561 aaaaaaaaaa aaaaaactat cagtaaaaca aaatacttaa tgtacttacc tcaatatgct
     4621 tccttaggtt tgatggcgaa cttttgaagg ccgatatttc tttattaagc gggagacaga
     4681 ggacacattt catcttgaat gaatctttat ttacaccact taaagaaaag aattcgcgaa
     4741 gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg
     4801 ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata atttccctaa
     4861 tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag
     4921 gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat
     4981 aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg
     5041 gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta
     5101 aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga
     5161 tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc
     5221 aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc
     5281 cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     5341 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5401 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5461 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5521 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5581 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5641 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5701 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5761 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5821 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5881 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5941 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6001 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6061 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6121 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6181 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6241 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6301 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6361 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6421 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7501 cgcgcacatt tccccg
//
