LOCUS       sequence-201376-       10794 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 gain-of-function allele of gpa-12 expressed 
            specifically in the adult epidermis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10794)
  TITLE     Ewbank lab plasmids
REFERENCE   2  (bases 1 to 10794)
  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"
COMMENT     Sequence Label: pNP148
FEATURES             Location/Qualifiers
     source          1..10794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(4134..4158)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4184..4200)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4184..4200)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4937..4955)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             7037..7048
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(8684..9272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8764..8783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9443..10234,10235..10303))
                     /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     10066..10085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10304..10408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10476..10494
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10532..10554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10654..10673
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
       61 cgccagcacg tgtgtttttg aacacggcga tatgtatctg tagatcatcc ccaattttca
      121 acaatgaatc taaatactga actttttgtg taatctttag tgcagaaaaa tgaccaaaat
      181 cagcgatttt ccgattatac ccaaaatatc tcagctcaat gagcacctat aaaaatgtaa
      241 aaataaagaa agtagtcgaa ccatattttg taaaggctaa aaaactgaat aaatttttat
      301 ccatccagcg cggcgttaca atccgtggcc catgtcacgc acggctttca aagaaaaaaa
      361 aacagtctgt ttgcgcgaca tcgatctggc aaaaatccta aaaacgacta tgcgtggctc
      421 actttatatt aatatgaaaa atacaaactt ggtggtgcag gtgctgcaga ggccatcgat
      481 gaagaagtgg aggccttgtt gtcgatctac aaaattgtat ggttgtttga gattttcctt
      541 gaaacgctct caatcttgga tatgatataa attctggata agatatgatg gatatgatct
      601 atgtttgata gagtgcattt gaaaggataa cggccctgca gacatttttg aaaaaataac
      661 aaacatttaa ttttctcact cgtttaggct attcccctat ttttgatatt ccttcgcaca
      721 tatgaaaact actttttttc gaaactgtta actccagaat ttataaatct atagccctta
      781 cttgattatt tattatcatg gttactcacc tcatgtctct ttctcttttc aagaagtctt
      841 cgttcatatc tgttgtctct ttcctccttt tgaatctctg ctttgctctt cgccattgtt
      901 tcctgaaaat aatgtaactt gaattgtgta atatactttt ttaatttgaa tttggcttgt
      961 aacgcggaat cactacgtgc gggatcattt cttactagaa aacccagaaa atgccatatt
     1021 tcacttatct cggggtcatt tctaattaga aaagctacaa aaaccatttc taattacggt
     1081 gactttagaa ctggtcgaaa tagctaatta gaaatgggtt tttgctaatt agaggtgact
     1141 ctatgttatc tgagtaagcc tcgtttgtat gtaaactcac agtactaata aatgcaagac
     1201 acccgggttt gtctagatat gaaataattg aaatatcaat tctgacagac aataatggta
     1261 atcttgataa ggagttccac gcccaggaga acacgttagt tttcttgttt ttgattgcgt
     1321 gcgttatttt ggagaaaaac tcgatttttt acaaaataat tttttgaaag gaacactgtt
     1381 caataagttt tgtctttttt ctcagttgtg atacggtttt ttattctttt ttgtagttat
     1441 acagaagacc gttacgaaac agactgatat cgaaacaaac actgatctta cttgcactta
     1501 taatacgact cactagtggg cagatcttcg aatggctccg tcatgtctga gctcaatttt
     1561 cctcattttt ctgatataaa ttgtcgaaaa ggtgtgccag actgctaacc agtttgacga
     1621 acaaaaaggc gggatgagca gtataataca gcttctgaat acaaagttca gggagagtac
     1681 ggaaagggat gacgtcacga gatcattctg aagataagat ggtaagatgg agattgtttt
     1741 taaaaatcaa aagtcgatca tctattcata gattgataac acggtcagaa tgatttgatg
     1801 tattcgattg ggtgatcttt gacacaaatt tgaacatacc aatacttgac aaaagtgtac
     1861 atacagtttt ttttttctag tattcgaagt tgtgtcaatt agatacaagt ttggaaaatt
     1921 ttgttgttgc cgaaattttt gcaaagcttc cttaatatga tacattttaa attaaactga
     1981 aagagaagaa aattgaattg gccagtcaat cttaaaaatg acttgcgtct cttcgggaac
     2041 gtataatatt ttttaatgaa catcgccaca aagaagtcaa cagattacag caaacttctc
     2101 tggaacccat tctcttcttg tccttttttc gaaactttac actgtgaacc taatctgaac
     2161 agttcaagtg atggctattg ctttaaaagg agtgactgaa aattgttgga acacagtcat
     2221 tcacctggta gaatggtatg ctgtttcggg aaaaaagatg aaaggaccaa aacgattgaa
     2281 aaagaattac acaaagagag aaaaattatg cggcgacaag tataattaaa ttattttcta
     2341 gtgcaaaata atcgacaatt ttcagattaa tttattgctt ctcggatccg gtgaatctgg
     2401 aaagtccacg tttgtaaaac agatgcacat aattcacggg gccggagaat ttacagcaga
     2461 cgaagttcga gcttacagac agcagatata tcaggtttca tccttttacg tggtttaaaa
     2521 aattcaaaga aaattattca cagaacgcga taagtgcaat gcgggtgttg ttagatgcta
     2581 ggaacaaatt aggaattgca tgggaagatc caaaaagaca agtggaagtt gaaaaagtta
     2641 tgaggtaagt agaatttata tctaaagtct gaatacaatc caaaattcta catttttcct
     2701 agagatcaag aaatgcgtga atcaaacatt caactgttat agttttaaga agaatttttg
     2761 gtatcgttta aaaataacaa agaactggat gaaaaatttt taacaatgaa aacgaaatta
     2821 gattattcgt tctaatcaaa attccacact ttttagaaat cattcaaaaa attaactttt
     2881 tttcttattg cacttaaata cattaatatt atagattttc tgtcggcgat ttattaaaag
     2941 gaattgattt caccacgttt gttgaagtag ccccaatcat cagtgatttc tggaacgacg
     3001 ctgcgattag aaaaacatat gagcaaagaa acttattcca aattgtaagt ttttgatttt
     3061 tttaaattat atatgttctg ttttcagagt gactcctgtc agtacttctt tgagcatatt
     3121 ccacgaatcg caatgcctga tttttaccca acaaaccgtg atatcctttt ctgccgaaaa
     3181 gcgacacgtg gcatttctga acacattttc gagatcaaca agattccatt taggtttgac
     3241 ttttttcctc aatggtcacg gacttagttt gtgaattata ttgtttcagg ttcattgatg
     3301 ttggcggaca aagatcgcaa cgtcaaaaat ggtttcaatg ttttacagac atcacaagta
     3361 ttttgttcat ggtcgccagt aacgaatacg atcaggttag agatcaagac acacttgttc
     3421 tggtgtttac tatacatttt tcaggtgatt ctcgaagaca gacggacgaa tcgagttgtc
     3481 gaatcacggt ctgtttttga gacgattgtc aacaacagag cgttctctaa tgtctccatc
     3541 atcttgttca tgaacaaaaa tgatttattg caagagaaag ttcccaaatc cgatatcaga
     3601 cagtatttca cggatttcac aggggatcac actttagtga gtctatatta ttgagaaaaa
     3661 acgtgtttga caatagaaaa atgtatcttg tatcaagcta tattaaaagg aaaatcatat
     3721 ttggtaaaaa ttaggggagt attacacagc cgacatgttt cgggcattgg aattcacgta
     3781 ttacggaaat ctttttttct ttattcattc tttcactgga actgtgaggg tacagttgga
     3841 aatgttgtgt tttactcacg accttttcaa atctcgaaaa atttcaataa ttaacgttaa
     3901 aagttttttt ttcttagagc aaaaaagttg ttatttaaaa acttgttctt ttccaggtcc
     3961 gagatgttca atttttcctt gtcgacaaat tcgaagcatc tcgccgcgat cgtgcacgac
     4021 ccttctttta tcacttcacc actgccgtcg acactgaaaa cattagaaga gtgttccgag
     4081 atgtacgtga aagtatttta gagcaaaact tgaagactct catgatgcag taagttcagc
     4141 tttcttgtac aaagtggtga tatcgaattc ctgcagcccg ggggatccac tagttctaga
     4201 gcggccgcca ccgcggccgc tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc
     4261 taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta
     4321 tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca
     4381 cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa
     4441 aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc
     4501 tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa
     4561 ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc
     4621 ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt
     4681 atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt
     4741 ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca
     4801 caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt
     4861 ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgccga
     4921 gctccagctt ttgttccctt tagtgagggt taattggata tctggatcca cgaagcttcc
     4981 catggtgacg tcaccggttc tagataccta ggtgagctct ggtaccctct agtcaaggcc
     5041 ttaagtgagt cgtattacgt agcccgggcc tagttctaga cattctctaa tgaaaaaatc
     5101 tttcagttga aattgaaaat gagttaaagt tggagttttt attgaaaaca gatttccgtg
     5161 tgattagtgt ttttagcgag tgtgacagga cagcgaaaaa atatagaaac aaggggggaa
     5221 ctgaaaagct taggaatgca ttgaacatga gaaggggaag gggaaggaac aaactagaca
     5281 ggaattattg gaatttaatc acatttggag ttttttttct attcgacaga ataattatcc
     5341 agaacatttt tgtattaaat atttatgcat catatgagta gtcggctttg ttgtgcatga
     5401 cgagtttgtt atcgacgaaa tagaagctgt cagaacgagt ctcgtttgga ttgttgatca
     5461 tgtcgtccac tgaaaaagag attagtcttt tgaattgtac tttttagaat aatgactcac
     5521 tgagttgttg agagagttga gggaactcat agatatgttc acagttgttt cgtgaattcg
     5581 gaatacagaa tccgaattca aagtcaaaac acttcagaag gcgatctttg aagaagtgac
     5641 gttcgatcat tcggaaatga tggattggga tgtctcctac tttgaattcc acagttgctc
     5701 cgaccgtttt gagtttcaaa aagtttggag caaatctata acgcacgtat cttgccgact
     5761 cctgtggcga ctcatcattc tcctgatcgt tctccggttt ggcgatctcg aaaagcactt
     5821 gctcagtgtc cagatcacgg atttggaact tggtgaactc gatgttatag atgttcgcag
     5881 atggggagca taagaatcct aaatttatgt tttaaactga aatccaaagg gagcaagata
     5941 ccttgagtga ttcccggaag tgctaaaacg tcgttcggag tgatttgagc tttcttcgca
     6001 agctccgatt ccgttgtgat tccttgttcg gtgcttggtg gtggccgtgg catctggaaa
     6061 tatggaaaag ttcaacaaaa agaaaagaga aaagaatgaa atcggatatc aagagttagt
     6121 tgagcggttt ctctagtttt ctgagtctca cctgcgacgg gaaggtcgcc gagccgggtg
     6181 gaatcgatcg ttgttgctcg gctttcatat cggtttggtt ggaagcggct gaaaacggaa
     6241 agaagtggaa gaaggaaaag agtgtggtgt gacaggaaaa tggtaattag agggtgccaa
     6301 ataaccagct atattttgtt tttttttgaa aacattttta aaaagaaaaa tacgataatg
     6361 atatcagatg gatttccgga aaactggtat gaaaaatttc aacctttttg agtacatgta
     6421 atcaaaatac actttgtaaa ttatcatttt tattgaaact ccaccatttt tctatttata
     6481 acgctaataa tttgaaaaag aaacctgttg cgaaccgcgg ggtgaatccc aaaaacgaat
     6541 gcgttttggt ggagtgattg attcgaatcg aagaagaaaa agaagaagac gtggaataga
     6601 gagctcactc ttaaccgagc agcacacacc gacagaaaaa aaaatgaaat gaatgagggt
     6661 cttcttcttc ttcttcttcg aatgattgac agaaatggga aaaagaggaa gattgagaag
     6721 ggaaaaagga aggagaaaag aagcagaaga agacgtcaga gaggagagga acgagcggaa
     6781 aagcagcggg cgcaagtcat agaagtagca gagctgggga gaagaagaca ctatccaaga
     6841 aaggaatgac gagagagtat gcaaaggggt atagggtgca gacagaatag gaacagaata
     6901 acagatgatg agccaagaag agttgaaaag ggcgatgaat ttgtcatgta acttaatttg
     6961 ggtcaatttg agcatgatga attgaaatca tcccttgttg ggagttaata accggtttgt
     7021 tatcagaaac cctgtaatag aagggcgccc taactttgag ccaattcatc ccggtttctg
     7081 tcaaatatat caaaaagtgg tcaactgaca aattgttttt gatattataa taaacatttt
     7141 atccgttaac aattttcgaa tactttttac aaggacttgg ataaattggc tcaaagacga
     7201 agtgagtttg ctaccatcat ggaatcagga ggacatttcg acaatgtcat ataatgtttg
     7261 aaaaatgatt ttaacaaggg tctcattcga gataagcaat gctctgagca agtggctcgt
     7321 cgtcttcaag aatctcttgt tgatattcat ttggatcctg ttttgcacga agcatgtgcc
     7381 atggatattc agagtttttg caaaaaagtt ccgccaggtc aaacaatctt tttaattttt
     7441 atatgatact tcgggtgtat ttttcttctt tttcctgttc attttcgctt ttttctcact
     7501 tttttgtaac aaaatctctc acttttctgt aagtcttaac aaaagttcaa ttggctcgat
     7561 caagtaaaat ttcggccgta tgaacgtttt caaaggtttt catttttttt taagttcatt
     7621 cacaatatca tgaatctctc gtcctttggt ttccggtaag tagcgaaata atataagcat
     7681 acttataaat gaaggaattg aatatagaat aaggaaagca taactaccaa taacggaata
     7741 tagaggtaat actgtaaatg tactgataac caccattaca gtattcattg aatatgccac
     7801 agattgagca atactacgat gtttttgagg taccagttca gaggatataa accacgaaat
     7861 tggaccaact ccagttctga aatttcatta ttcgattaaa tcaaaatata tttgaaaaac
     7921 ctcacccata agtataaccg tagatgatca atgcaactag gcatccgtat ttcatcgggt
     7981 caatcagaat acgtattttt gcgaagaata cgaatgcaat tagagctaaa ttattgagaa
     8041 atgtgaagag aaggagaagc ttgcgtcgac caactctgta aacttattca tacaattttg
     8101 tcttcaaatt ttgaacgttt aaaactatct tattcaatga gttagcgggt ttttatagtg
     8161 agttagttca cgattatcta ataaaactat gtccaaacat cctaactgcg accgtattct
     8221 cactaactcg tcttaaaaac gttgagatta ttttaaaccc ttcttcagta caactacagt
     8281 actgatgcac aaatacaata ataataatac attcagacat actctgaaca atgtaacttg
     8341 tttttgttat actgtggtaa catttattga aatgaaatat aaccttactt ttcaatcaca
     8401 attccaccag taattgttcc agaaacataa gctaaaccca ttgcagttgt actccattca
     8461 gcaatttctg actcaacatt tgcattttct agaaagaaag tagaggataa aagaagagcc
     8521 cataatccga cagggccctg tgaaattgtt atccgctggt atcagctcac tcaaaggcgg
     8581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8641 agcaaaagac caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     9001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     9061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     9121 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     9181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     9241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     9301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    10021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    10081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    10141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    10201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    10321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    10381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
    10441 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
    10501 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
    10561 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
    10621 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
    10681 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
    10741 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcct
//
