LOCUS       sequence-204633-        9438 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 9438)
  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 9438)
  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..9438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             615..665
                     /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"
     primer_bind     3795..3817
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3855..3873)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3941..4045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4046..4114,4115..4906)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4264..4283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5077..5665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5566..5585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7301..7312)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gggatgagca gtataataca gcttctgaat acaaagttca gggagagtac ggaaagggat
       61 gacgtcacga gatcattctg aagataagat ggtaagatgg agattgtttt taaaaatcaa
      121 aagtcgatca tctattcata gattgataac acggtcagaa tgatttgatg tattcgattg
      181 ggtgatcttt gacacaaatt tgaacatacc aatacttgac aaaagtgtac atacagtttt
      241 ttttttctag tattcgaagt tgtgtcaatt agatacaagt ttggaaaatt ttgttgttgc
      301 cgaaattttt gcaaagcttc cttaatatga tacattttaa attaaactga aagagaagaa
      361 aattgaattg gccagtcaat cttaaaaatg acttgcgtct cttcgggaac gtataatatt
      421 ttttaatgaa catcgccaca aagaagtcaa cagattacag caaacttctc tggaacccat
      481 tctcttcttg tccttttttc gaaactttac actgtgaacc taatctgaac agttcaagtg
      541 atggctattg ctttaaaagg agtgactgaa aattgttgga acacagtcat tcacctggta
      601 gaatgagcgg ccgcatggga gttgctgatc ttattaaaaa attcgaatct atttctaaag
      661 aagaaggagg aggaggcgcg ccaagcgctg ctggttccat gtccgagctc atcaaggaga
      721 acatgcacat gaagctctac atggagggaa ccgtcaacaa ccaccacttc aagtgcacct
      781 ccgagggaga gggaaagcca tacgagggaa cccaaaccat gcgtatcaag gtaagtttaa
      841 acatatatat actaactaac cctgattatt taaattttca ggccgtcgag ggaggaccac
      901 tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatccaag accttcatca
      961 accacaccca aggaatccca gacttcttca agcaatcctt cccagaggga ttcacctggg
     1021 agcgtgtcac cacctacgag gacggaggag tcctcaccgc cacccaagac acctccctcc
     1081 aagacggatg cctcatctac aacgtcaagg taagtttaaa cagttcggta ctaactaacc
     1141 atacatattt aaattttcag atccgtggag tcaacttccc atccaacgga ccagtcatgc
     1201 aaaagaagac cctcggatgg gaggcctcca ccgagaccct ctacccagcc gacggaggac
     1261 tcgagggacg tgccgacatg gccctcaagc tcgtcggagg aggacacctc atctgcaacc
     1321 tcaaggtaag tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagacca
     1381 cctaccgttc caagaagcca gccaagaacc tcaagatgcc aggagtctac tacgtcgacc
     1441 gtcgtctcga gcgtatcaag gaggccgaca aggagaccta cgtcgagcaa cacgaggtcg
     1501 ccgtcgcccg ttactgcgac ctcccatcca agctcggaca ccgttaagcc ggccctctaa
     1561 agtttttaat tccggattat ttttttgttt aatttactga cctgattgta tccaaacgtt
     1621 cccaattctt atccattccc ctatgttttt agaccatgat tgtccccttg atcaacattt
     1681 ccctgcacaa tctgctttaa tagtattcgc cgtgttataa atttgcacaa tatcgcatgt
     1741 cacgtcgttc ttctcgtttc atgtctatca ccatcatcac ctcagcttca atatgtattt
     1801 ttatcattta ccgcttcccg gtagaacatt tttcgtacgg atagtgtccc aaataatgcc
     1861 ccaacattgc ttcccattcc ccaggcacta tccggttgtc ccaagtttaa attattatcg
     1921 agttgaagat gacaaataaa aacgaaattt gtgttttcga gtaaattgaa gacgctcatt
     1981 ttactgcaaa caaccgctat cagatctgcc cactagtgag tcgtattata agtgcaagta
     2041 agatcagtgt ttgtttcgat atcagtctgt ttcgtaacgg tcttctgtat aactacaaaa
     2101 aagaataaaa aaccgtatca caactgagaa aaaagacaaa acttattgaa cagtgttcct
     2161 ttcaaaaaat tattttgtaa aaaatcgagt ttttctccaa aataacgcac gcaatcaaaa
     2221 acaagaaaac taacgtgttc tcctgggcgt ggaactcctt atcaagatta ccattattgt
     2281 ctgtcagaat tgatatttca attatttcat atctagacaa acccgggtgt cttgcattta
     2341 ttagtactgt gagtttacat acaaacgagg cttactcaga taacatagag tcacctctaa
     2401 ttagcaaaaa cccatttcta attagctatt tcgaccagtt ctaaagtcac cgtaattaga
     2461 aatggttttt gtagcttttc taattagaaa tgaccccgag ataagtgaaa tatggcattt
     2521 tctgggtttt ctagtaagaa atgatcccgc acgtagtgat tccgcgttac aagccaaatt
     2581 caaattaaaa aagtatatta cacaattcaa gttacattat tttcaggaaa caatggcgaa
     2641 gagcaaagca gagattcaaa aggaggaaag agacaacaga tatgaacgaa gacttcttga
     2701 aaagagaaag agacatgagg tgagtaacca tgataataaa taatcaagta agggctatag
     2761 atttataaat tctggagtta acagtttcga aaaaaagtag ttttcatatg tgcgaaggaa
     2821 tatcaaaaat aggggaatag cctaaacgag tgagaaaatt aaatgtttgt tattttttca
     2881 aaaatgtctg cagggccgtt atcctttcaa atgcactcta tcaaacatag atcatatcca
     2941 tcatatctta tccagaattt atatcatatc caagattgag agcgtttcaa ggaaaatctc
     3001 aaacaaccat acaattttgt agatcgacaa caaggcctcc acttcttcat cgatggcctc
     3061 tgcagcacct gcaccaccaa gtttgtattt ttcatattaa tataaagtga gccacgcata
     3121 gtcgttttta ggatttttgc cagatcgatg tcgcgcaaac agactgtttt tttttctttg
     3181 aaagccgtgc gtgacatggg ccacggattg taacgccgcg ctggatggat aaaaatttat
     3241 tcagtttttt agcctttaca aaatatggtt cgactacttt ctttattttt acatttttat
     3301 aggtgctcat tgagctgaga tattttgggt ataatcggaa aatcgctgat tttggtcatt
     3361 tttctgcact aaagattaca caaaaagttc agtatttaga ttcattgttg aaaattgggg
     3421 atgatctaca gatacatatc gccgtgttca aaaacacacg tgctgtgacg tcgtgactgg
     3481 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3541 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3601 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3661 atggtgcact ctcagtgcaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3721 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3781 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3841 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3901 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3961 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4021 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4081 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4141 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4201 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4261 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4321 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4381 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4441 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4501 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4561 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4621 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4681 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggt cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt
     5701 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5761 ccgtattacc gcctttgagt gagctgatac cagcggataa caatttcaca gggccctgtc
     5821 ggattatggg ctcttctttt atcctctact ttctttctag aaaatgcaaa tgttgagtca
     5881 gaaattgctg aatggagtac aactgcaatg ggtttagctt atgtttctgg aacaattact
     5941 ggtggaattg tgattgaaaa gtaaggttat atttcatttc aataaatgtt accacagtat
     6001 aacaaaaaca agttacattg ttcagagtat gtctgaatgt attattatta ttgtatttgt
     6061 gcatcagtac tgtagttgta ctgaagaagg gtttaaaata atctcaacgt ttttaagacg
     6121 agttagtgag aatacggtcg cagttaggat gtttggacat agttttatta gataatcgtg
     6181 aactaactca ctataaaaac ccgctaactc attgaataag atagttttaa acgttcaaaa
     6241 tttgaagaca aaattgtatg aataagttta cagagttggt cgacgcaagc ttctccttct
     6301 cttcacattt ctcaataatt tagctctaat tgcattcgta ttcttcgcaa aaatacgtat
     6361 tctgattgac ccgatgaaat acggatgcct agttgcattg atcatctacg gttatactta
     6421 tgggtgaggt ttttcaaata tattttgatt taatcgaata atgaaatttc agaactggag
     6481 ttggtccaat ttcgtggttt atatcctctg aactggtacc tcaaaaacat cgtagtattg
     6541 ctcaatctgt ggcatattca atgaatactg taatggtggt tatcagtaca tttacagtat
     6601 tacctctata ttccgttatt ggtagttatg ctttccttat tctatattca attccttcat
     6661 ttataagtat gcttatatta tttcgctact taccggaaac caaaggacga gagattcatg
     6721 atattgtgaa tgaacttaaa aaaaaatgaa aacctttgaa aacgttcata cggccgaaat
     6781 tttacttgat cgagccaatt gaacttttgt taagacttac agaaaagtga gagattttgt
     6841 tacaaaaaag tgagaaaaaa gcgaaaatga acaggaaaaa gaagaaaaat acacccgaag
     6901 tatcatataa aaattaaaaa gattgtttga cctggcggaa cttttttgca aaaactctga
     6961 atatccatgg cacatgcttc gtgcaaaaca ggatccaaat gaatatcaac aagagattct
     7021 tgaagacgac gagccacttg ctcagagcat tgcttatctc gaatgagacc cttgttaaaa
     7081 tcatttttca aacattatat gacattgtcg aaatgtcctc ctgattccat gatggtagca
     7141 aactcacttc gtctttgagc caatttatcc aagtccttgt aaaaagtatt cgaaaattgt
     7201 taacggataa aatgtttatt ataatatcaa aaacaatttg tcagttgacc actttttgat
     7261 atatttgaca gaaaccggga tgaattggct caaagttagg gcgcccttct attacagggt
     7321 ttctgataac aaaccggtta ttaactccca acaagggatg atttcaattc atcatgctca
     7381 aattgaccca aattaagtta catgacaaat tcatcgccct tttcaactct tcttggctca
     7441 tcatctgtta ttctgttcct attctgtctg caccctatac ccctttgcat actctctcgt
     7501 cattcctttc ttggatagtg tcttcttctc cccagctctg ctacttctat gacttgcgcc
     7561 cgctgctttt ccgctcgttc ctctcctctc tgacgtcttc ttctgcttct tttctccttc
     7621 ctttttccct tctcaatctt cctctttttc ccatttctgt caatcattcg aagaagaaga
     7681 agaagaagac cctcattcat ttcatttttt tttctgtcgg tgtgtgctgc tcggttaaga
     7741 gtgagctctc tattccacgt cttcttcttt ttcttcttcg attcgaatca atcactccac
     7801 caaaacgcat tcgtttttgg gattcacccc gcggttcgca ataggtttct ttttcaaatt
     7861 attagcgtta taaatagaaa aatggtggag tttcaataaa aatgataatt tacaaagtgt
     7921 attttgatta catgtactca aaaaggttga aatttttcat accagttttc cggaaatcca
     7981 tctgatatca ttatcgtatt tttcttttta aaaatgtttt caaaaaaaaa caaaatatag
     8041 ctggttattt ggcaccctct aattaccatt ttcctgtcac accacactct tttccttctt
     8101 ccacttcttt ccgttttcag ccgcttccaa ccaaaccgat atgaaagccg agcaacaacg
     8161 atcgattcca cccggctcgg cgaccttccc gtcgcaggtg agactcagaa aactagagaa
     8221 accgctcaac taactcttga tatccgattt cattcttttc tcttttcttt ttgttgaact
     8281 tttccatatt tccagatgcc acggccacca ccaagcaccg aacaaggaat cacaacggaa
     8341 tcggagcttg cgaagaaagc tcaaatcact ccgaacgacg ttttagcact tccgggaatc
     8401 actcaaggta tcttgctccc tttggatttc agtttaaaac ataaatttag gattcttatg
     8461 ctccccatct gcgaacatct ataacatcga gttcaccaag ttccaaatcc gtgatctgga
     8521 cactgagcaa gtgcttttcg agatcgccaa accggagaac gatcaggaga atgatgagtc
     8581 gccacaggag tcggcaagat acgtgcgtta tagatttgct ccaaactttt tgaaactcaa
     8641 aacggtcgga gcaactgtgg aattcaaagt aggagacatc ccaatccatc atttccgaat
     8701 gatcgaacgt cacttcttca aagatcgcct tctgaagtgt tttgactttg aattcggatt
     8761 ctgtattccg aattcacgaa acaactgtga acatatctat gagttccctc aactctctca
     8821 acaactcagt gagtcattat tctaaaaagt acaattcaaa agactaatct ctttttcagt
     8881 ggacgacatg atcaacaatc caaacgagac tcgttctgac agcttctatt tcgtcgataa
     8941 caaactcgtc atgcacaaca aagccgacta ctcatatgat gcataaatat ttaatacaaa
     9001 aatgttctgg ataattattc tgtcgaatag aaaaaaaact ccaaatgtga ttaaattcca
     9061 ataattcctg tctagtttgt tccttcccct tccccttctc atgttcaatg cattcctaag
     9121 cttttcagtt ccccccttgt ttctatattt tttcgctgtc ctgtcacact cgctaaaaac
     9181 actaatcaca cggaaatctg ttttcaataa aaactccaac tttaactcat tttcaatttc
     9241 aactgaaaga ttttttcatt agagaatgtc tagaactagg cccgggctac gtaatacgac
     9301 tcacttaagg ccttgactag agggtaccag agctcaccta gggctccgtc atgtctgagc
     9361 tcaattttcc tcatttttct gatataaatt gtcgaaaagg tgtgccagac tgctaaccag
     9421 tttgacgaac aaaaaggc
//
