LOCUS       sequence-227236-        5960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PXC1 J2_pECIA14 should be used with bait vector PXC1 
            J2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  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 5960)
  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: PXC1 J2_pECIA14
FEATURES             Location/Qualifiers
     source          1..5960
                     /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             1256..1279
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2954..2977
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2993..3010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3033..3050)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3083..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3107..3126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3161..3180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3361..3377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3361..3377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3374..3396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3385..3401
                     /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(3409..3415,3416..3433,3434..3439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3454..3475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3592..3609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3763..4351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3843..3862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..5313,5314..5382))
                     /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     5145..5164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5383..5487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5555..5573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5611..5633)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5733..5752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5946..5960,1..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 aacgacacaa atgctctcac
      541 actcttccgt ctccaaaccg acacacacgg caacctcgcc ggaaactgga ccggctccga
      601 cgcttgtacc tcctcatggc aaggcgtttc ttgctcacca tcttcacacc gcgtcactga
      661 actatctctc ccttccctct ccttacgagg accattaacc tctctctcct ccttagacca
      721 acttcgtctt cttgacctcc acgataacag actcaacggc accgtttcac cactcacaaa
      781 ctgtaaaaac ctcagactcg tttatctcgc cggtaacgat ctctccggtg agattcctaa
      841 agagatatcc tttcttaaac ggatgatacg tcttgatctc tctgataaca atatccgtgg
      901 agttatacca agagagattc tcgggtttac ccgggtttta acgatccgga tccagaacaa
      961 tgagttaacg ggtcggatcc cggatttttc tcagatgaag tcacttcttg aattgaatgt
     1021 ttccttcaat gagttgcacg gaaacgtctc cgacggcgtg gtgaagaaat ttggagactt
     1081 gagcttttcc ggtaacgaag gattatgcgg gtcggatcct ttaccggttt gtactatcac
     1141 aaacgacccg gaatcttcta acactgatca gattgttccg tcgaatccta cttcgattcc
     1201 gcattctccg gttagcgtta gagaaccgga aattcacagt catagaggga ttaaactgga
     1261 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1321 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1381 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1441 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1501 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1561 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     1621 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     1681 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     1741 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     1801 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     1861 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     1921 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     1981 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     2041 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2101 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2161 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2221 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2281 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2341 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2401 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2461 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2521 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     2581 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     2641 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     2701 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     2761 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     2821 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     2881 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     2941 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     3001 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3061 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3121 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3181 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3241 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3301 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5521 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5581 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5641 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5701 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5761 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5821 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5881 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5941 gttttcccag tcacgacgtt
//
