LOCUS       sequence-270292-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector IMK2 J9_pECIA14 should be used with bait vector IMK2 
            J9_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  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 6476)
  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: IMK2 J9_pECIA14
FEATURES             Location/Qualifiers
     source          1..6476
                     /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             1772..1795
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3470..3493
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3509..3526
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3549..3566)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3599..3733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3623..3642)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3677..3696
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3877..3893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3877..3893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3890..3912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3901..3917
                     /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(3925..3931,3932..3949,3950..3955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3970..3991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4108..4125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4359..4378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5038..5829,5830..5898))
                     /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     5661..5680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5899..6003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6071..6089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6127..6149)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6249..6268
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6462..6476,1..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 ggtggtggtc actcttggga
      541 tggaatcgtg gtgactcagg cgaactatca ggcactccaa gctatcaaac acgaactcat
      601 tgacttcacc ggagttctca aaagctggaa caactctgcc tcctctcaag tttgctccgg
      661 ctgggctgga atcaaatgcc tccgaggtca agtggtcgcc attcagctcc cttggaaggg
      721 acttggcggt actatctctg agaagattgg acagcttgga agtctcagaa aacttagcct
      781 ccacaacaat gttatcgccg gttcggttcc tcgttccctt ggctacctca agagcctccg
      841 tggagtctat ctcttcaaca atcgtctctc gggttccatc ccagtctcac tgggtaactg
      901 ccctcttctc cagaatcttg atctcagcag taaccagcta actggagcca tcccgcctag
      961 ccttactgaa tccacgaggc tctataggct caatctcagc ttcaattcac tttccggtcc
     1021 ccttccggtt agtgtagcca gatcatatac cctcactttt cttgatcttc agcataacaa
     1081 cctatctggt tcgatacctg actttttcgt caatggctcc caccctctca aaacgctgaa
     1141 tcttgaccac aatcgcttct ctggagctgt tccagtgtct ctctgcaagc atagtctgtt
     1201 ggaagaggtt tctataagcc ataaccagct ctcaggttct attcccagag aatgtggagg
     1261 tcttccacac ctccagagcc tggatttctc ttacaattca atcaacggaa ccatcccaga
     1321 cagtttctcc aacctttcat ctctagtttc actaaatctc gaaagcaacc acctcaaagg
     1381 cccaatccca gatgccattg atagacttca caacctgaca gagcttaacc tgaagagaaa
     1441 caagatcaat gggccaatcc cagaaacaat agggaacata tctggaatca aaaagcttga
     1501 tctgtcagaa aacaacttca caggtccaat cccgctctca ctagtccacc tggcgaagct
     1561 ttcttcattc aacgtctcct acaacaccct ctctggtcct gttccacctg tcctctccaa
     1621 gaagttcaac tcaagctctt tcttgggcaa catccagctt tgtggctaca gctcctcaaa
     1681 tccatgccct gctcctgatc accaccatcc cctcaccctt tcacctacct catcacaaga
     1741 accaagaaaa caccaccaca gaaaactctc actggaggtg ctgttccagg gaccaagcgg
     1801 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     1861 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     1921 caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg gctccatcat
     1981 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     2041 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     2101 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     2161 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2221 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2281 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2341 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2401 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     2461 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     2521 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     2581 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     2641 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     2701 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     2761 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     2821 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     2881 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     2941 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     3001 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     3061 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     3121 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3181 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3241 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3301 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3361 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3421 caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg attacaagga
     3481 tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag tttaaacccg
     3541 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     3601 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3661 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3721 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3781 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3841 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3901 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3961 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4021 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5101 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5341 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5641 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5881 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5941 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6001 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6061 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6121 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6181 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6241 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6301 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6361 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6421 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
