LOCUS       sequence-298977-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G62710 X045_pECIA2 should be used with prey vector 
            AT5G62710 X045_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4913)
  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 4913)
  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: AT5G62710 X045_pECIA2
FEATURES             Location/Qualifiers
     source          1..4913
                     /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             1166..1189
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1895..1936
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1946..1963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1986..2003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2036..2170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2114..2133
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2314..2330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2314..2330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2327..2349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2338..2354
                     /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(2362..2368,2369..2386,2387..2392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2407..2428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2545..2562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2796..2815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3475..4266,4267..4335))
                     /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     4098..4117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4336..4440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4508..4526
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4564..4586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4686..4705
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4899..4913,1..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 tcctttgctc tcactcttga
      541 tggatttgct ctattggaat tgaagagtgg atttaatgat acgaggaact cactagagaa
      601 ctggaaagat tcagatgagt ctccttgttc ttggactggt gtctcctgta atcctcaaga
      661 ccaaagagtt gtttctataa acttaccata tatgcaatta ggagggataa tatctcctag
      721 cattgggaag cttagtcgat tgcagagact ggcacttcat cagaatagct tacatgggaa
      781 cattcctaat gaaatcacca actgcactga gctaagagct atgtacttga gggcgaattt
      841 tctccaaggc gggattccac cggatttagg caaccttaca tttcttacta tattggatct
      901 atcaagcaat acactcaagg gtgctattcc ttcttcaatt agtcgattga cgcggttacg
      961 ctccttgaac ttgtcaacca actttttctc tggtgaaata ccggatattg gagttctcag
     1021 cagattcggg gtcgaaactt tcactggtaa tttggatctc tgtggccggc aaattcgcaa
     1081 gccatgtaga tcatcaatgg gtttccctgt tgttcttcct catgcagaaa gtgctgatga
     1141 atcagattct ccaaagcgat catcactgga ggtgctgttc cagggaccag ggagcgacaa
     1201 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     1261 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     1321 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     1381 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     1441 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     1501 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     1561 gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga ccaagaacca
     1621 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     1681 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     1741 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     1801 cttctcatgc tccgtgatgc atgagggtct gcacaaccac tacacgcaga agagcctctc
     1861 cctgtctccg ggtaaatcta gagggccctt cgaaggtaag cctatcccta accctctcct
     1921 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     1981 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     2041 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2101 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2161 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2221 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     2281 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4441 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4501 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4561 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4621 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4681 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4741 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     4801 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4861 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
