LOCUS       sequence-296809-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector IMK2 J9_pECIA2 should be used with prey vector IMK2 
            J9_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  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 5519)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..5519
                     /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             2501..2542
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2552..2569
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2592..2609)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2642..2776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2666..2685)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2720..2739
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2920..2936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2920..2936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2933..2955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2944..2960
                     /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(2968..2974,2975..2992,2993..2998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3013..3034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3151..3168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3322..3910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3402..3421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4081..4872,4873..4941))
                     /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     4704..4723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4942..5046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5114..5132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5170..5192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5292..5311
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5505..5519,1..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 gaccagggag
     1801 cgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg gaccgtcagt
     1861 cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac
     1921 atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga
     1981 cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta
     2041 ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca aggagtacaa
     2101 gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct ccaaagccaa
     2161 agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg agatgaccaa
     2221 gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca tcgccgtgga
     2281 gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc
     2341 cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt ggcagcaggg
     2401 gaacgtcttc tcatgctccg tgatgcatga gggtctgcac aaccactaca cgcagaagag
     2461 cctctccctg tctccgggta aatctagagg gcccttcgaa ggtaagccta tccctaaccc
     2521 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     2581 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     2641 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2701 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2761 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2821 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     2881 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4201 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4441 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4921 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4981 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5041 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5101 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5161 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5221 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5281 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5341 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5461 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
