LOCUS       sequence-297043-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G01950 X064_pECIA2 should be used with prey vector 
            AT5G01950 X064_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  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 5843)
  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: AT5G01950 X064_pECIA2
FEATURES             Location/Qualifiers
     source          1..5843
                     /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             2096..2119
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2825..2866
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2876..2893
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2916..2933)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2966..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2990..3009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3044..3063
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3244..3260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3244..3260)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3257..3279)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3268..3284
                     /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(3292..3298,3299..3316,3317..3322))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3337..3358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3475..3492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3646..4234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3726..3745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4405..5196,5197..5265))
                     /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     5028..5047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5266..5370)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5438..5456
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5494..5516)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5616..5635
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5829..5843,1..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 acccaccctt cagaagtcac
      541 tgctttacga tctgttaaaa gaagtctact tgaccctaaa gattatttga gaaactggaa
      601 tagaggcgac ccttgcagat caaattggac tggagttatc tgctttaatg agattggtac
      661 tgacgactac ctgcatgtcc gagaactcct gctgatgaat atgaatctct ccgggacttt
      721 gtcaccagag ctccaaaaat tagctcatct cgaaatatta gatttcatgt ggaataatat
      781 aagtggttca attccaaacg agataggaca gatatcatcc ctggtactct tgctgctgaa
      841 tggaaacaaa ctttcaggga ctttaccaag cgaactcggg tatctttcga acttgaacag
      901 gttccagata gatgagaata acatcacagg accaatacca aagtcctttt ccaacttgaa
      961 aaaagtgaaa catcttcact tcaacaataa ttcgttgacc ggccagattc cagttgagct
     1021 ttccaatctt actaacatct ttcatgtgct gctagacaac aataaattat ctggaaatct
     1081 tccaccacaa ctctctgcat tgccaaacct gcaaatcctg caacttgata ataacaactt
     1141 cagtggatcc gatatcccag cttcctatgg aaacttttcg aatattctaa agctgagtct
     1201 tagaaattgt agtttgaaag gggcacttcc tgattttagc aagatacgtc atctcaaata
     1261 cttggatctt agctggaatg aactcacagg acctatacct tcctctaatt tttctaagga
     1321 tgtgacaacc attaatttat caaacaacat acttaacgga tccatcccgc agagcttctc
     1381 agatctccct ttacttcaga tgctattgct gaaaaacaat atgttatctg gatctgtccc
     1441 tgattccctt tggaagaata tctcttttcc gaagaaagct agacttctac ttgatcttag
     1501 gaataattct ctgtcacgtg ttcaaggaga tctaactcct ccacaaaatg tcactttaag
     1561 gcttgatggc aatctgatct gcactaatgg aagcatatca aacgcaaacc tattctgtga
     1621 gtccaaaggc aaagagtgga tatcactacc aaacaattca accaattctg cactggactg
     1681 tcctcctctt gcctgtccca ctcctgactt ctacgaatac agtccagcat ccccattacg
     1741 atgcttttgt gcagcgcctc ttagaattgg atatcgtctc aaaagtccta gtttctcata
     1801 cttccctcca tatattgatc aatttggcga gtatgttacc gatttccttc aaatggaacc
     1861 atatcaactc tggatcgact catatcaatg ggagaaaggg cctcgtctga ggatgtatct
     1921 gaaacttttt ccgaaagtta atgaaaccta tacaagaaca tttaacgaga gtgaagttct
     1981 cagaatccga ggcatattcg cttcatggag atttcctggt agtgacttat ttggcccgta
     2041 tgaactgctc aacttcactc tgcaggggcc ttattcatat gttaacttca acagtctgga
     2101 ggtgctgttc cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     2161 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     2221 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     2281 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     2341 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     2401 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     2461 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     2521 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     2581 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     2641 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     2701 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     2761 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     2821 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     2881 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     2941 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     3001 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3061 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3121 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3181 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     3241 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3301 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4321 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4381 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4501 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4561 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4921 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5281 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5341 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5401 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5461 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5581 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5641 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     5701 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5761 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5821 agggttttcc cagtcacgac gtt
//
