LOCUS       sequence-201377-        9412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To insert a single copy of a construct at a site on Chr II, 
            corresponding to a mKate-labelled LifeAct expressed specifically in 
            the adult epidermis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9412)
  AUTHORS   Taffoni C, Omi S, Huber C, Mailfert S, Fallet M, Rupprecht JF, 
            Ewbank JJ, Pujol N
  TITLE     Microtubule plus-end dynamics link wound repair to the innate immune
            response.
  JOURNAL   Elife. 2020 Jan 29;9. pii: 45047. doi: 10.7554/eLife.45047.
  PUBMED    31995031
REFERENCE   2  (bases 1 to 9412)
  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"; journalName: "Elife."; 
            date: "2020-01-29"; volume: "9. pii: 45047. doi"; pages: " 
            10.7554/eLife.45047"
COMMENT     Sequence Label: pNP151
FEATURES             Location/Qualifiers
     source          1..9412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(99..687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(179..198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(858..1649,1650..1718))
                     /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     1481..1500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1719..1823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1891..1909
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1947..1969)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2069..2088
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5073..5123)
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             7864..7875
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
       61 aagaccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
      541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1861 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     1921 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1981 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2041 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2101 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     2161 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2221 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagcacgt
     2281 gtgtttttga acacggcgat atgtatctgt agatcatccc caattttcaa caatgaatct
     2341 aaatactgaa ctttttgtgt aatctttagt gcagaaaaat gaccaaaatc agcgattttc
     2401 cgattatacc caaaatatct cagctcaatg agcacctata aaaatgtaaa aataaagaaa
     2461 gtagtcgaac catattttgt aaaggctaaa aaactgaata aatttttatc catccagcgc
     2521 ggcgttacaa tccgtggccc atgtcacgca cggctttcaa agaaaaaaaa acagtctgtt
     2581 tgcgcgacat cgatctggca aaaatcctaa aaacgactat gcgtggctca ctttatatta
     2641 atatgaaaaa tacaaacttg gtggtgcagg tgctgcagag gccatcgatg aagaagtgga
     2701 ggccttgttg tcgatctaca aaattgtatg gttgtttgag attttccttg aaacgctctc
     2761 aatcttggat atgatataaa ttctggataa gatatgatgg atatgatcta tgtttgatag
     2821 agtgcatttg aaaggataac ggccctgcag acatttttga aaaaataaca aacatttaat
     2881 tttctcactc gtttaggcta ttcccctatt tttgatattc cttcgcacat atgaaaacta
     2941 ctttttttcg aaactgttaa ctccagaatt tataaatcta tagcccttac ttgattattt
     3001 attatcatgg ttactcacct catgtctctt tctcttttca agaagtcttc gttcatatct
     3061 gttgtctctt tcctcctttt gaatctctgc tttgctcttc gccattgttt cctgaaaata
     3121 atgtaacttg aattgtgtaa tatacttttt taatttgaat ttggcttgta acgcggaatc
     3181 actacgtgcg ggatcatttc ttactagaaa acccagaaaa tgccatattt cacttatctc
     3241 ggggtcattt ctaattagaa aagctacaaa aaccatttct aattacggtg actttagaac
     3301 tggtcgaaat agctaattag aaatgggttt ttgctaatta gaggtgactc tatgttatct
     3361 gagtaagcct cgtttgtatg taaactcaca gtactaataa atgcaagaca cccgggtttg
     3421 tctagatatg aaataattga aatatcaatt ctgacagaca ataatggtaa tcttgataag
     3481 gagttccacg cccaggagaa cacgttagtt ttcttgtttt tgattgcgtg cgttattttg
     3541 gagaaaaact cgatttttta caaaataatt ttttgaaagg aacactgttc aataagtttt
     3601 gtcttttttc tcagttgtga tacggttttt tattcttttt tgtagttata cagaagaccg
     3661 ttacgaaaca gactgatatc gaaacaaaca ctgatcttac ttgcacttat aatacgactc
     3721 actagtgggc agatctgata gcggttgttt gcagtaaaat gagcgtcttc aatttactcg
     3781 aaaacacaaa tttcgttttt atttgtcatc ttcaactcga taataattta aacttgggac
     3841 aaccggatag tgcctgggga atgggaagca atgttggggc attatttggg acactatccg
     3901 tacgaaaaat gttctaccgg gaagcggtaa atgataaaaa tacatattga agctgaggtg
     3961 atgatggtga tagacatgaa acgagaagaa cgacgtgaca tgcgatattg tgcaaattta
     4021 taacacggcg aatactatta aagcagattg tgcagggaaa tgttgatcaa ggggacaatc
     4081 atggtctaaa aacatagggg aatggataag aattgggaac gtttggatac aatcaggtca
     4141 gtaaattaaa caaaaaaata atccggaatt aaaaacttta gagggccggc ttaacggtgt
     4201 ccgagcttgg atgggaggtc gcagtaacgg gcgacggcga cctcgtgttg ctcgacgtag
     4261 gtctccttgt cggcctcctt gatacgctcg agacgacggt cgacgtagta gactcctggc
     4321 atcttgaggt tcttggctgg cttcttggaa cggtaggtgg tctgaaaatt taaatcagat
     4381 tagttagtta gtaaaatcat gtttaaactt accttgaggt tgcagatgag gtgtcctcct
     4441 ccgacgagct tgagggccat gtcggcacgt ccctcgagtc ctccgtcggc tgggtagagg
     4501 gtctcggtgg aggcctccca tccgagggtc ttcttttgca tgactggtcc gttggatggg
     4561 aagttgactc cacggatctg aaaatttaaa tatgtatggt tagttagtac cgaactgttt
     4621 aaacttacct tgacgttgta gatgaggcat ccgtcttgga gggaggtgtc ttgggtggcg
     4681 gtgaggactc ctccgtcctc gtaggtggtg acacgctccc aggtgaatcc ctctgggaag
     4741 gattgcttga agaagtctgg gattccttgg gtgtggttga tgaaggtctt ggatccgtac
     4801 atgaaggagg tggcgaggat gtcgaaggcg aatgggagtg gtcctccctc gacggcctga
     4861 aaatttaaat aatcagggtt agttagtata tatatgttta aacttacctt gatacgcatg
     4921 gtttgggttc cctcgtatgg ctttccctct ccctcggagg tgcacttgaa gtggtggttg
     4981 ttgacggttc cctccatgta gagcttcatg tgcatgttct ccttgatgag ctcggacatg
     5041 gaaccagcag cgcttggcgc gcctcctcct ccttcttctt tagaaataga ttcgaatttt
     5101 ttaataagat cagcaactcc catgcggccg ctcattctac caggtgaatg actgtgttcc
     5161 aacaattttc agtcactcct tttaaagcaa tagccatcac ttgaactgtt cagattaggt
     5221 tcacagtgta aagtttcgaa aaaaggacaa gaagagaatg ggttccagag aagtttgctg
     5281 taatctgttg acttctttgt ggcgatgttc attaaaaaat attatacgtt cccgaagaga
     5341 cgcaagtcat ttttaagatt gactggccaa ttcaattttc ttctctttca gtttaattta
     5401 aaatgtatca tattaaggaa gctttgcaaa aatttcggca acaacaaaat tttccaaact
     5461 tgtatctaat tgacacaact tcgaatacta gaaaaaaaaa actgtatgta cacttttgtc
     5521 aagtattggt atgttcaaat ttgtgtcaaa gatcacccaa tcgaatacat caaatcattc
     5581 tgaccgtgtt atcaatctat gaatagatga tcgacttttg atttttaaaa acaatctcca
     5641 tcttaccatc ttatcttcag aatgatctcg tgacgtcatc cctttccgta ctctccctga
     5701 actttgtatt cagaagctgt attatactgc tcatcccgcc tttttgttcg tcaaactggt
     5761 tagcagtctg gcacaccttt tcgacaattt atatcagaaa aatgaggaaa attgagctca
     5821 gacatgacgg agccctaggt gagctctggt accctctagt caaggcctta agtgagtcgt
     5881 attacgtagc ccgggcctag ttctagacat tctctaatga aaaaatcttt cagttgaaat
     5941 tgaaaatgag ttaaagttgg agtttttatt gaaaacagat ttccgtgtga ttagtgtttt
     6001 tagcgagtgt gacaggacag cgaaaaaata tagaaacaag gggggaactg aaaagcttag
     6061 gaatgcattg aacatgagaa ggggaagggg aaggaacaaa ctagacagga attattggaa
     6121 tttaatcaca tttggagttt tttttctatt cgacagaata attatccaga acatttttgt
     6181 attaaatatt tatgcatcat atgagtagtc ggctttgttg tgcatgacga gtttgttatc
     6241 gacgaaatag aagctgtcag aacgagtctc gtttggattg ttgatcatgt cgtccactga
     6301 aaaagagatt agtcttttga attgtacttt ttagaataat gactcactga gttgttgaga
     6361 gagttgaggg aactcataga tatgttcaca gttgtttcgt gaattcggaa tacagaatcc
     6421 gaattcaaag tcaaaacact tcagaaggcg atctttgaag aagtgacgtt cgatcattcg
     6481 gaaatgatgg attgggatgt ctcctacttt gaattccaca gttgctccga ccgttttgag
     6541 tttcaaaaag tttggagcaa atctataacg cacgtatctt gccgactcct gtggcgactc
     6601 atcattctcc tgatcgttct ccggtttggc gatctcgaaa agcacttgct cagtgtccag
     6661 atcacggatt tggaacttgg tgaactcgat gttatagatg ttcgcagatg gggagcataa
     6721 gaatcctaaa tttatgtttt aaactgaaat ccaaagggag caagatacct tgagtgattc
     6781 ccggaagtgc taaaacgtcg ttcggagtga tttgagcttt cttcgcaagc tccgattccg
     6841 ttgtgattcc ttgttcggtg cttggtggtg gccgtggcat ctggaaatat ggaaaagttc
     6901 aacaaaaaga aaagagaaaa gaatgaaatc ggatatcaag agttagttga gcggtttctc
     6961 tagttttctg agtctcacct gcgacgggaa ggtcgccgag ccgggtggaa tcgatcgttg
     7021 ttgctcggct ttcatatcgg tttggttgga agcggctgaa aacggaaaga agtggaagaa
     7081 ggaaaagagt gtggtgtgac aggaaaatgg taattagagg gtgccaaata accagctata
     7141 ttttgttttt ttttgaaaac atttttaaaa agaaaaatac gataatgata tcagatggat
     7201 ttccggaaaa ctggtatgaa aaatttcaac ctttttgagt acatgtaatc aaaatacact
     7261 ttgtaaatta tcatttttat tgaaactcca ccatttttct atttataacg ctaataattt
     7321 gaaaaagaaa cctgttgcga accgcggggt gaatcccaaa aacgaatgcg ttttggtgga
     7381 gtgattgatt cgaatcgaag aagaaaaaga agaagacgtg gaatagagag ctcactctta
     7441 accgagcagc acacaccgac agaaaaaaaa atgaaatgaa tgagggtctt cttcttcttc
     7501 ttcttcgaat gattgacaga aatgggaaaa agaggaagat tgagaaggga aaaaggaagg
     7561 agaaaagaag cagaagaaga cgtcagagag gagaggaacg agcggaaaag cagcgggcgc
     7621 aagtcataga agtagcagag ctggggagaa gaagacacta tccaagaaag gaatgacgag
     7681 agagtatgca aaggggtata gggtgcagac agaataggaa cagaataaca gatgatgagc
     7741 caagaagagt tgaaaagggc gatgaatttg tcatgtaact taatttgggt caatttgagc
     7801 atgatgaatt gaaatcatcc cttgttggga gttaataacc ggtttgttat cagaaaccct
     7861 gtaatagaag ggcgccctaa ctttgagcca attcatcccg gtttctgtca aatatatcaa
     7921 aaagtggtca actgacaaat tgtttttgat attataataa acattttatc cgttaacaat
     7981 tttcgaatac tttttacaag gacttggata aattggctca aagacgaagt gagtttgcta
     8041 ccatcatgga atcaggagga catttcgaca atgtcatata atgtttgaaa aatgatttta
     8101 acaagggtct cattcgagat aagcaatgct ctgagcaagt ggctcgtcgt cttcaagaat
     8161 ctcttgttga tattcatttg gatcctgttt tgcacgaagc atgtgccatg gatattcaga
     8221 gtttttgcaa aaaagttccg ccaggtcaaa caatcttttt aatttttata tgatacttcg
     8281 ggtgtatttt tcttcttttt cctgttcatt ttcgcttttt tctcactttt ttgtaacaaa
     8341 atctctcact tttctgtaag tcttaacaaa agttcaattg gctcgatcaa gtaaaatttc
     8401 ggccgtatga acgttttcaa aggttttcat ttttttttaa gttcattcac aatatcatga
     8461 atctctcgtc ctttggtttc cggtaagtag cgaaataata taagcatact tataaatgaa
     8521 ggaattgaat atagaataag gaaagcataa ctaccaataa cggaatatag aggtaatact
     8581 gtaaatgtac tgataaccac cattacagta ttcattgaat atgccacaga ttgagcaata
     8641 ctacgatgtt tttgaggtac cagttcagag gatataaacc acgaaattgg accaactcca
     8701 gttctgaaat ttcattattc gattaaatca aaatatattt gaaaaacctc acccataagt
     8761 ataaccgtag atgatcaatg caactaggca tccgtatttc atcgggtcaa tcagaatacg
     8821 tatttttgcg aagaatacga atgcaattag agctaaatta ttgagaaatg tgaagagaag
     8881 gagaagcttg cgtcgaccaa ctctgtaaac ttattcatac aattttgtct tcaaattttg
     8941 aacgtttaaa actatcttat tcaatgagtt agcgggtttt tatagtgagt tagttcacga
     9001 ttatctaata aaactatgtc caaacatcct aactgcgacc gtattctcac taactcgtct
     9061 taaaaacgtt gagattattt taaacccttc ttcagtacaa ctacagtact gatgcacaaa
     9121 tacaataata ataatacatt cagacatact ctgaacaatg taacttgttt ttgttatact
     9181 gtggtaacat ttattgaaat gaaatataac cttacttttc aatcacaatt ccaccagtaa
     9241 ttgttccaga aacataagct aaacccattg cagttgtact ccattcagca atttctgact
     9301 caacatttgc attttctaga aagaaagtag aggataaaag aagagcccat aatccgacag
     9361 ggccctgtga aattgttatc cgctggtatc agctcactca aaggcggtaa ta
//
