LOCUS       sequence-296845-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G45340 K10_pECIA2 should be used with prey vector 
            AT2G45340 K10_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  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 5126)
  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: AT2G45340 K10_pECIA2
FEATURES             Location/Qualifiers
     source          1..5126
                     /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             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2108..2149
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2159..2176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2199..2216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2249..2383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2273..2292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2327..2346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2527..2543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2527..2543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2540..2562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2551..2567
                     /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(2575..2581,2582..2599,2600..2605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2758..2775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2929..3517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3009..3028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3688..4479,4480..4548))
                     /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     4311..4330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4549..4653)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4721..4739
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4777..4799)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4899..4918
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5112..5126,1..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 tcctcctcag agctcgacat
      541 tctcctcgat atcaaatcct ctctcgaccc tgaaaaacgt ttcctcactt catggactcc
      601 cgacgccgac ccttgctcct ctggctcctt cgacggtgtc gcgtgcgacg gaaaccgccg
      661 tgttgctaac atctctcttc aaggaatggg tctcactgga actatccctc cttcaatcgg
      721 tcttctcaca agcttaaccg gtttatacct tcacttcaac tccttgaccg gtcatattcc
      781 taaagatatc tcaaatctgc ctctcctcac cgatttatac ctcaatgtca acaatctctc
      841 cggtgaaatc cctcctctaa tcgggaactt agacaatctt caagttatac agttatgtta
      901 caacaagtta agtggaagta taccgacaca gtttggttct cttaagaaga taactgtctt
      961 ggctttacag tataaccaac tctccggcgc cattccggcg agtttaggcg atattgatac
     1021 gttaacgagg cttgatttga gcttcaacaa cctctttggt cctgttccgg tgaaactcgc
     1081 cggtgctcct ttgcttgaag ttctcgacat tcgtaataac tccttctccg gcttcgttcc
     1141 ctctgctttg aagaggctaa acaatggatt ccagtattca aacaatcacg gtttatgcgg
     1201 agatggtttc actgatttga aagcttgcac cggcttaaac ggtccgaacc ctaaccgacc
     1261 agacccaaca aacccaacga atttcacaac agttgacgtg aaaccagaat cagcagattt
     1321 acaacgaagc aattgcagta acaacaatgg cgggtgttca tcaaaatcct taaagtctct
     1381 ggaggtgctg ttccagggac cagggagcga caaaactcac acatgcccac cgtgcccagc
     1441 acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct
     1501 catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc
     1561 tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc
     1621 gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca
     1681 ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc
     1741 catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg tgtacaccct
     1801 gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg
     1861 cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta
     1921 caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac
     1981 cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggg
     2041 tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaat ctagagggcc
     2101 cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     2161 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     2221 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     2281 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2341 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2401 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2461 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     2521 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2581 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2641 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2701 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2761 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2821 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3121 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3361 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3601 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3661 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3721 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3781 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3841 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3901 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3961 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4021 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4081 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4141 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4201 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4261 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4321 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4381 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4441 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4501 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4561 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4621 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4681 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4741 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4801 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4861 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     4921 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4981 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5041 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5101 gccagggttt tcccagtcac gacgtt
//
