LOCUS       sequence-296817-        4943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G24100 K4_pECIA2 should be used with prey vector 
            AT5G24100 K4_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4943)
  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 4943)
  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: AT5G24100 K4_pECIA2
FEATURES             Location/Qualifiers
     source          1..4943
                     /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             1196..1219
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1925..1966
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1976..1993
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2016..2033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2066..2200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2090..2109)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2144..2163
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2344..2360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2344..2360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2357..2379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2368..2384
                     /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(2392..2398,2399..2416,2417..2422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2437..2458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2575..2592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3505..4296,4297..4365))
                     /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     4128..4147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4366..4470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4538..4556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4594..4616)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4716..4735
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4929..4943,1..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 tctcaggtca ccggtgatct
      541 agccggtgac agacaagccc ttctcgattt cctcaacaac atcattcacc cacggtctct
      601 agcgtggaac acaagcagcc cggtatgcac cacgtggcca ggggtcacat gcgacataga
      661 cggcacacgt gtcactgccc ttcacttgcc cggagcaagt cttctaggag tgatcccacc
      721 tggtactatc agccgtctat cggagcttca gatcctcagt cttcgatcca acggcctacg
      781 agggccgttc ccaattgatt tcttgcagct taagaaactc aaagcaatta gtctcgggaa
      841 caacagattc tctggcccat taccatccga ttacgccacg tggacgaatc tcactgttct
      901 tgatctgtac agtaaccggt ttaacggtag tattcccgcg ggatttgcta atcttaccgg
      961 actcgtctcg cttaacctag ctaaaaactc attctccggt gagattccgg atcttaatct
     1021 ccccggtcta cgcagactta acttttccaa caacaatctc accggatcaa tcccaaattc
     1081 tctgaaaaga ttcgggaact cggctttttc cggcaacaac ttagtcttcg aaaatgcccc
     1141 tcctccggcg gtagtttcat tcaaagagca aaagaaaaac ggcatctata tctctctgga
     1201 ggtgctgttc cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     1261 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     1321 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     1381 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     1441 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     1501 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     1561 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     1621 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     1681 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     1741 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     1801 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     1861 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     1921 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     1981 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     2041 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     2101 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2161 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2221 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2281 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     2341 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2401 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2461 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4561 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4621 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4681 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4741 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4801 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4861 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4921 agggttttcc cagtcacgac gtt
//
