LOCUS       sequence-296774-        4883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G64210 I9_pECIA2 should be used with prey vector 
            AT1G64210 I9_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4883)
  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 4883)
  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: AT1G64210 I9_pECIA2
FEATURES             Location/Qualifiers
     source          1..4883
                     /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             1136..1159
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1865..1906
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1916..1933
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1956..1973)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2006..2140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2030..2049)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2084..2103
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2284..2300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2284..2300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2297..2319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2308..2324
                     /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(2332..2338,2339..2356,2357..2362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2377..2398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2515..2532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2686..3274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2766..2785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3445..4236,4237..4305))
                     /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     4068..4087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4306..4410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4478..4496
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4534..4556)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4656..4675
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4869..4883,1..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 tctcagactc ttgaagacga
      541 caagaaagct ttgcttcatt tcttgtctag tttcaattct tctcgtctcc attggaacca
      601 gagctcagac gtttgtcata gctggaccgg tgttacctgc aacgaaaacg gagatcgaat
      661 tgtatccgtt agattgcctg cggttggatt caacggtttg attcctcctt tcaccatcag
      721 tcgtctctct tctctcaagt ttctaagcct tagaaagaac catttcaccg gagacttccc
      781 ttctgatttc acaaatctca agagcttgac tcatctctac ctacaacata accacctctc
      841 aggtccacta ctagctatct tctcggagtt gaagaatctt aaggttcttg atctgtctaa
      901 caatggattc aacggaagca tcccaacctc tctttcagga ttgactagcc ttcaagtctt
      961 gaatcttgca aacaattcat tctcagggga aattccaaat cttcatcttc caaaactgag
     1021 tcagatcaac ttgtccaaca acaagctcat cggtacaata ccaaagtctc ttcaaagatt
     1081 ccaaagttca gctttttctg gcaacaacct taccgagagg aagaaacaac gcaagctgga
     1141 ggtgctgttc cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     1201 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     1261 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     1321 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     1381 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     1441 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     1501 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     1561 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     1621 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     1681 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     1741 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     1801 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     1861 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     1921 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     1981 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     2041 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2101 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2161 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2221 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     2281 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2341 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2401 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2461 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2521 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2581 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2641 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2701 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2761 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2821 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2881 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2941 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3001 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3061 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3121 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3181 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3241 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3301 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3361 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3421 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3481 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3541 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3601 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3661 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3721 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3781 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3841 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3901 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3961 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4021 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4081 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4141 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4201 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4261 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4381 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4441 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4501 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4561 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4621 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4681 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4741 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4801 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4861 agggttttcc cagtcacgac gtt
//
