LOCUS       sequence-204634-       10821 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 10821)
  TITLE     Ewbank lab plasmids
REFERENCE   2  (bases 1 to 10821)
  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..10821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(102..893,894..962))
                     /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     725..744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(963..1067)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1135..1153
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1191..1213)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    complement(5614..5638)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5664..5680)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5664..5680)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(6417..6435)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             8517..8528
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(10164..10752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10244..10263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
       61 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      181 gtgtagataa ctacgatacg ggagggctta ccatctgacc ccagtgctgc aatgataccg
      241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
      421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
      481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
      541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
      601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
      661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
      721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
      781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
      841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
      901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1081 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1141 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     1201 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     1261 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     1321 gagcagattg cactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     1381 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1441 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1501 ttaagttggg taacgccagg gttttcccca gtcacgacgt cacagcacgt gtgtttttga
     1561 acacggcgat atgtatctgt agatcatccc caattttcaa caatgaatct aaatactgaa
     1621 ctttttgtgt aatctttagt gcagaaaaat gaccaaaatc agcgattttc cgattatacc
     1681 caaaatatct cagctcaatg agcacctata aaaatgtaaa aataaagaaa gtagtcgaac
     1741 catattttgt aaaggctaaa aaactgaata aatttttatc catccagcgc ggcgttacaa
     1801 tccgtggccc atgtcacgca cggctttcaa agaaaaaaaa acagtctgtt tgcgcgacat
     1861 cgatctggca aaaatcctaa aaacgactat gcgtggctca ctttatatta atatgaaaaa
     1921 tacaaacttg gtggtgcagg tgctgcagag gccatcgatg aagaagtgga ggccttgttg
     1981 tcgatctaca aaattgtatg gttgtttgag attttccttg aaacgctctc aatcttggat
     2041 atgatataaa ttctggataa gatatgatgg atatgatcta tgtttgatag agtgcatttg
     2101 aaaggataac ggccctgcag acatttttga aaaaataaca aacatttaat tttctcactc
     2161 gtttaggcta ttcccctatt tttgatattc cttcgcacat atgaaaacta ctttttttcg
     2221 aaactgttaa ctccagaatt tataaatcta tagcccttac ttgattattt attatcatgg
     2281 ttactcacct catgtctctt tctcttttca agaagtcttc gttcatatct gttgtctctt
     2341 tcctcctttt gaatctctgc tttgctcttc gccattgttt cctgaaaata atgtaacttg
     2401 aattgtgtaa tatacttttt taatttgaat ttggcttgta acgcggaatc actacgtgcg
     2461 ggatcatttc ttactagaaa acccagaaaa tgccatattt cacttatctc ggggtcattt
     2521 ctaattagaa aagctacaaa aaccatttct aattacggtg actttagaac tggtcgaaat
     2581 agctaattag aaatgggttt ttgctaatta gaggtgactc tatgttatct gagtaagcct
     2641 cgtttgtatg taaactcaca gtactaataa atgcaagaca cccgggtttg tctagatatg
     2701 aaataattga aatatcaatt ctgacagaca ataatggtaa tcttgataag gagttccacg
     2761 cccaggagaa cacgttagtt ttcttgtttt tgattgcgtg cgttattttg gagaaaaact
     2821 cgatttttta caaaataatt ttttgaaagg aacactgttc aataagtttt gtcttttttc
     2881 tcagttgtga tacggttttt tattcttttt tgtagttata cagaagaccg ttacgaaaca
     2941 gactgatatc gaaacaaaca ctgatcttac ttgcacttat aatacgactc acgagtgggc
     3001 agatcttcga atggctccgt catgtctgag ctcaattttc ctcatttttc tgatataaat
     3061 tgtcgaaaag gtgtgccaga ctgctaacca gtttgacgaa caaaaaggcg ggatgagcag
     3121 tataatacag cttctgaata caaagttcag ggagagtacg gaaagggatg acgtcacgag
     3181 atcattctga agataagatg gtaagatgga gattgttttt aaaaatcaaa agtcgatcat
     3241 ctattcatag attgataaca cggtcagaat gatttgatgt attcgattgg gtgatctttg
     3301 acacaaattt gaacatacca atacttgaca aaagtgtaca tacagttttt tttttctagt
     3361 attcgaagtt gtgtcaatta gatacaagtt tggaaaattt tgttgttgcc gaaatttttg
     3421 caaagcttcc ttaatatgat acattttaaa ttaaactgaa agagaagaaa attgaattgg
     3481 ccagtcaatc ttaaaaatga cttgcgtctc ttcgggaacg tataatattt tttaatgaac
     3541 atcgccacaa agaagtcaac agattacagc aaacttctct ggaacccatt ctcttcttgt
     3601 ccttttttcg aaactttaca ctgtgaacct aatctgaaca gttcaagtga tggctattgc
     3661 tttaaaagga gtgactgaaa attgttggaa cacagtcatt cacctggtag aatggtatgc
     3721 tgtttcggga aaaaagatga aaggaccaaa acgattgaaa aagaattaca caaagagaga
     3781 aaaattatgc ggcgacaagt ataattaaat tattttctag tgcaaaataa tcgacaattt
     3841 tcagattaat ttattgcttc tcggatccgg tgaatctgga aagtccacgt ttgtaaaaca
     3901 gatgcacata attcacgggg ccggagaatt tacagcagac gaagttcgag cttacagaca
     3961 gcagatatat caggtttcat ccttttacgt ggtttaaaaa attcaaagaa aattattcac
     4021 agaacgcgat aagtgcaatg cgggtgttgt tagatgctag gaacaaatta ggaattgcat
     4081 gggaagatcc aaaaagacaa gtggaagttg aaaaagttat gaggtaagta gaatttatat
     4141 ctaaagtctg aatacaatcc aaaattctac atttttccta gagatcaaga aatgcgtgaa
     4201 tcaaacattc aactgttata gttttaagaa gaatttttgg tatcgtttaa aaataacaaa
     4261 gaactggatg aaaaattttt aacaatgaaa acgaaattag attattcgtt ctaatcaaaa
     4321 ttccacactt tttagaaatc attcaaaaaa ttaacttttt ttcttattgc acttaaatac
     4381 attaatatta tagattttct gtcggcgatt tattaaaagg aattgatttc accacgtttg
     4441 ttgaagtagc cccaatcatc agtgatttct ggaacgacgc tgcgattaga aaaacatatg
     4501 agcaaagaaa cttattccaa attgtaagtt tttgattttt ttaaattata tatgttctgt
     4561 tttcagagtg actcctgtca gtacttcttt gagcatattc cacgaatcgc aatgcctgat
     4621 ttttacccaa caaaccgtga tatccttttc tgccgaaaag cgacacgtgg catttctgaa
     4681 cacattttcg agatcaacaa gattccattt aggtttgact tttttcctca atggtcacgg
     4741 acttagtttg tgaattatat tgtttcaggt tcattgatgt tggcggacta agatcgcaac
     4801 gtcaaaaatg gtttcaatgt tttacagaca tcacaagtat tttgttcatg gtcgccagta
     4861 acgaatacga tcaggttaga gatcaagaca cacttgttct ggtgtttact atacattttt
     4921 caggtgattc tcgaagacag acggacgaat cgagttgtcg aatcacggtc tgtttttgag
     4981 acgattgtca acaacagagc gttctctaat gtctccatca tcttgttcat gaacaaaaat
     5041 gatttattgc aagagaaagt tcccaaatcc gatatcagac agtatttcac ggatttcaca
     5101 ggggatcaca ctttagtgag tctatattat tgagaaaaaa cgtgtttgac aatagaaaaa
     5161 tgtatcttgt atcaagctat attaaaagga aaatcatatt tggtaaaaat taggggagta
     5221 ttacacagcc gacatgtttc gggcattgga attcacgtat tacggaaatc tttttttctt
     5281 tattcattct ttcactggaa ctgtgagggt acagttggaa atgttgtgtt ttactcacga
     5341 ccttttcaaa tctcgaaaaa tttcaataat taacgttaaa agtttttttt tcttagagca
     5401 aaaaagttgt tatttaaaaa cttgttcttt tccaggtccg agatgttcaa tttttccttg
     5461 tcgacaaatt cgaagcatct cgccgcgatc gtgcacgacc cttcttttat cacttcacca
     5521 ctgccgtcga cactgaaaac attagaagag tgttccgaga tgtacgtgaa agtattttag
     5581 agcaaaactt gaagactctc atgatgcagt aatacccagc tttcttgtac aaagtggtga
     5641 tatcgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggccgc
     5701 tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa
     5761 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     5821 cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg
     5881 tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc
     5941 ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt
     6001 tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat
     6061 catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc
     6121 atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc
     6181 aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt
     6241 tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     6301 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     6361 taatttgaaa gtggagtagt gtctatgggg tttttgccga gctccagctt ttgttccctt
     6421 tagtgagggt taattggata tctggatcca cgaagcttcc catggtgacg tcaccggttc
     6481 tagataccta ggtgagctct ggtaccctct agtcaaggcc ttaagtgagt cgtattacgt
     6541 agcccgggcc tagttctaga cattctctaa tgaaaaaatc tttcagttga aattgaaaat
     6601 gagttaaagt tggagttttt attgaaaaca gatttccgtg tgattagtgt ttttagcgag
     6661 tgtgacagga cagcgaaaaa atatagaaac aaggggggaa ctgaaaagct taggaatgca
     6721 ttgaacatga gaaggggaag gggaaggaac aaactagaca ggaattattg gaatttaatc
     6781 acatttggag ttttttttct attcgacaga ataattatcc agaacatttt tgtattaaat
     6841 atttatgcat catatgagta gtcggctttg ttgtgcatga cgagtttgtt atcgacgaaa
     6901 tagaagctgt cagaacgagt ctcgtttgga ttgttgatca tgtcgtccac tgaaaaagag
     6961 attagtcttt tgaattgtac tttttagaat aatgactcac tgagttgttg agagagttga
     7021 gggaactcat agatatgttc acagttgttt cgtgaattcg gaatacagaa tccgaattca
     7081 aagtcaaaac acttcagaag gcgatctttg aagaagtgac gttcgatcat tcggaaatga
     7141 tggattggga tgtctcctac tttgaattcc acagttgctc cgaccgtttt gagtttcaaa
     7201 aagtttggag caaatctata acgcacgtat cttgccgact cctgtggcga ctcatcattc
     7261 tcctgatcgt tctccggttt ggcgatctcg aaaagcactt gctcagtgtc cagatcacgg
     7321 atttggaact tggtgaactc gatgttatag atgttcgcag atggggagca taagaatcct
     7381 aaatttatgt tttaaactga aatccaaagg gagcaagata ccttgagtga ttcccggaag
     7441 tgctaaaacg tcgttcggag tgatttgagc tttcttcgca agctccgatt ccgttgtgat
     7501 tccttgttcg gtgcttggtg gtggccgtgg catctggaaa tatggaaaag ttcaacaaaa
     7561 agaaaagaga aaagaatgaa atcggatatc aagagttagt tgagcggttt ctctagtttt
     7621 ctgagtctca cctgcgacgg gaaggtcgcc gagccgggtg gaatcgatcg ttgttgctcg
     7681 gctttcatat cggtttggtt ggaagcggct gaaaacggaa agaagtggaa gaaggaaaag
     7741 agtgtggtgt gacaggaaaa tggtaattag agggtgccaa ataaccagct atattttgtt
     7801 tttttttgaa aacattttta aaaagaaaaa tacgataatg atatcagatg gatttccgga
     7861 aaactggtat gaaaaatttc aacctttttg agtacatgta atcaaaatac actttgtaaa
     7921 ttatcatttt tattgaaact ccaccatttt tctatttata acgctaataa tttgaaaaag
     7981 aaacctattg cgaaccgcgg ggtgaatccc aaaaacgaat gcgttttggt ggagtgattg
     8041 attcgaatcg aagaagaaaa agaagaagac gtggaataga gagctcactc ttaaccgagc
     8101 agcacacacc gacagaaaaa aaaatgaaat gaatgagggt cttcttcttc ttcttcttcg
     8161 aatgattgac agaaatggga aaaagaggaa gattgagaag ggaaaaagga aggagaaaag
     8221 aagcagaaga agacgtcaga gaggagagga acgagcggaa aagcagcggg cgcaagtcat
     8281 agaagtagca gagctgggga gaagaagaca ctatccaaga aaggaatgac gagagagtat
     8341 gcaaaggggt atagggtgca gacagaatag gaacagaata acagatgatg agccaagaag
     8401 agttgaaaag ggcgatgaat ttgtcatgta acttaatttg ggtcaatttg agcatgatga
     8461 attgaaatca tcccttgttg ggagttaata accggtttgt tatcagaaac cctgtaatag
     8521 aagggcgccc taactttgag ccaattcatc ccggtttctg tcaaatatat caaaaagtgg
     8581 tcaactgaca aattgttttt gatattataa taaacatttt atccgttaac aattttcgaa
     8641 tactttttac aaggacttgg ataaattggc tcaaagacga agtgagtttg ctaccatcat
     8701 ggaatcagga ggacatttcg acaatgtcat ataatgtttg aaaaatgatt ttaacaaggg
     8761 tctcattcga gataagcaat gctctgagca agtggctcgt cgtcttcaag aatctcttgt
     8821 tgatattcat ttggatcctg ttttgcacga agcatgtgcc atggatattc agagtttttg
     8881 caaaaaagtt ccgccaggtc aaacaatctt tttaattttt atatgatact tcgggtgtat
     8941 ttttcttctt tttcctgttc attttcgctt ttttctcact tttttgtaac aaaatctctc
     9001 acttttctgt aagtcttaac aaaagttcaa ttggctcgat caagtaaaat ttcggccgta
     9061 tgaacgtttt caaaggtttt catttttttt taagttcatt cacaatatca tgaatctctc
     9121 gtcctttggt ttccggtaag tagcgaaata atataagcat acttataaat gaaggaattg
     9181 aatatagaat aaggaaagca taactaccaa taacggaata tagaggtaat actgtaaatg
     9241 tactgataac caccattaca gtattcattg aatatgccac agattgagca atactacgat
     9301 gtttttgagg taccagttca gaggatataa accacgaaat tggaccaact ccagttctga
     9361 aatttcatta ttcgattaaa tcaaaatata tttgaaaaac ctcacccata agtataaccg
     9421 tagatgatca atgcaactag gcatccgtat ttcatcgggt caatcagaat acgtattttt
     9481 gcgaagaata cgaatgcaat tagagctaaa ttattgagaa atgtgaagag aaggagaagc
     9541 ttgcgtcgac caactctgta aacttattca tacaattttg tcttcaaatt ttgaacgttt
     9601 aaaactatct tattcaatga gttagcgggt ttttatagtg agttagttca cgattatcta
     9661 ataaaactat gtccaaacat cctaactgcg accgtattct cactaactcg tcttaaaaac
     9721 gttgagatta ttttaaaccc ttcttcagta caactacagt actgatgcac aaatacaata
     9781 ataataatac attcagacat actctgaaca atgtaacttg tttttgttat actgtggtaa
     9841 catttattga aatgaaatat aaccttactt ttcaatcaca attccaccag taattgttcc
     9901 agaaacataa gctaaaccca ttgcagttgt actccattca gcaatttctg actcaacatt
     9961 tgcattttct agaaagaaag tagaggataa aagaagagcc cataatccga cagggccctg
    10021 tgaaattgtt atccgctggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    10081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaagac caggaaccgt
    10141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    10201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    10261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    10321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    10381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    10441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    10501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    10561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    10621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    10681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    10741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    10801 aactcacgtt aagggatttt g
//
