LOCUS       sequence-270268-        5885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PRK2A I12_pECIA14 should be used with bait vector PRK2A 
            I12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5885)
  AUTHORS   Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, 
            Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger 
            N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, 
            Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir 
            Y
  TITLE     An extracellular network of Arabidopsis leucine-rich repeat receptor
            kinases.
  JOURNAL   Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. 
            Epub 2018 Jan 10.
  PUBMED    29320478
REFERENCE   2  (bases 1 to 5885)
  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: 
            "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 
            18"; volume: "553"; issue: "7688"; pages: "342-346"
COMMENT     Sequence Label: PRK2A I12_pECIA14
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        23..449
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     467..520
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1181..1204
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2879..2902
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2918..2935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2958..2975)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3008..3142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3032..3051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3086..3105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3286..3302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3286..3302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3299..3321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3310..3326
                     /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(3334..3340,3341..3358,3359..3364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3379..3400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4447..5238,5239..5307))
                     /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     5070..5089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5308..5412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5480..5498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5536..5558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5658..5677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5871..5885,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg tcagagaccg aaactctctt
      541 gaaattcaag aattctcttg ttattgggag agcaaatgct ttggagagct ggaatagaag
      601 gaatcctcca tgcaaatgga ctggtgtctt gtgtgataga ggctttgttt gggggttacg
      661 actagagaat cttgagctct ctggctctat cgacatcgag gcgttgatgg gtttgaattc
      721 tctaaggtct cttagtttca taaacaacaa atttaaaggt ccatttccag agttcaagaa
      781 acttgttgct ttgaaatcac tttacttgtc aaacaatcaa ttcgacctag agatacctaa
      841 ggatgcattt gatggcatgg ggtggttgaa aaaacttcat ctggagcaaa ataattttat
      901 tggtgaaatt ccaacgtcct tggtcaaaag tccaaagctt atagagctta gacttgacgg
      961 gaaccggttc accggacaaa taccagaatt cagacatcat ccaaacatgt taaacctttc
     1021 aaacaatgca ttggcaggtc aaatcccaaa tagtttcagt acaatggatc caaaattgtt
     1081 tgaagggaat aaaggcttat gtggaaaacc tttggataca aagtgttctt ctccttacaa
     1141 ccattcctcg gaacccaagt cgagcaccaa gaaaacatca ctggaggtgc tgttccaggg
     1201 accaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     1261 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     1321 cctgaagaac accgtgatgg agtgcgacgc ctgcggcatg cagccctcta gaggctccgg
     1381 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     1441 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     1501 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     1561 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     1621 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     1681 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     1741 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     1801 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     1861 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     1921 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     1981 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     2041 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     2101 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     2161 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     2221 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     2281 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     2341 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     2401 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     2461 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     2521 tggcaaggcc cgggacagga aggcctatac ggtcctccta tacggaaacg gtccaggcta
     2581 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     2641 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     2701 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     2761 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     2821 cgccggcacc accgacgccg cgcacccggg ttctagagga agcacctccg gcgataacga
     2881 ttacaaggat gatgatgata aggccagcgg caccggtcat catcaccatc accattgagt
     2941 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     3001 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3061 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3121 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3181 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3241 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3361 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5461 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5521 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5581 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5641 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5701 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5761 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5821 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5881 acgtt
//
