LOCUS       sequence-227296-        5003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector PXC1 J2_pECIA2 should be used with prey vector PXC1 
            J2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5003)
  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 5003)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..5003
                     /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             1985..2026
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2036..2053
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2076..2093)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2126..2260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2150..2169)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2204..2223
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2404..2420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2404..2420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2417..2439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2428..2444
                     /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(2452..2458,2459..2476,2477..2482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2497..2518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2635..2652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2806..3394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2886..2905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3565..4356,4357..4425))
                     /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     4188..4207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4426..4530)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4598..4616
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4654..4676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4776..4795
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4989..5003,1..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 cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     1321 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     1381 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     1441 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     1501 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     1561 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     1621 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     1681 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     1741 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     1801 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     1861 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     1921 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     1981 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     2041 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     2101 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     2161 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2221 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2281 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2341 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     2401 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2461 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2521 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2581 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2641 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2701 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2761 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2821 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2881 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2941 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3001 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3061 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3121 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3181 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3241 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3301 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3361 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3421 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3481 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3541 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3601 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3661 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3721 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3781 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3841 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3901 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3961 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4021 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4081 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4141 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4201 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4261 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4321 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4381 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4441 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4501 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4561 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4621 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4681 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4741 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4801 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4861 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4921 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4981 agggttttcc cagtcacgac gtt
//
