LOCUS       sequence-298813-        6416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G33170 X049_pECIA2 should be used with prey vector 
            AT2G33170 X049_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6416)
  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 6416)
  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: AT2G33170 X049_pECIA2
FEATURES             Location/Qualifiers
     source          1..6416
                     /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             2669..2692
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3398..3439
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3449..3466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3489..3506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3539..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3563..3582)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3617..3636
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3817..3833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3817..3833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3830..3852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3841..3857
                     /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(3865..3871,3872..3889,3890..3895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3910..3931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4048..4065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4219..4807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4299..4318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4978..5769,5770..5838))
                     /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     5601..5620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5839..5943)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6011..6029
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6067..6089)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6189..6208
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6402..6416,1..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 agtttgaata gtgatggtca
      541 gtttctcttg gagttgaaga acagaggatt tcaagattcg ttgaaccgtt tacataattg
      601 gaacggcatt gacgagacgc cgtgtaattg gataggtgtg aattgcagct cacaaggcag
      661 tagtagtagt agtaatagtc tggtggtgac atctcttgat ttgagttcta tgaatctctc
      721 tgggatagta agtcctagca ttggtggttt agttaacttg gtctatctta acctcgcgta
      781 caacgcgtta accggagata ttcctcggga gattggaaac tgttcgaagc tggaggttat
      841 gtttttgaac aataaccagt ttggtggttc cattccagtc gagatcaaca agttatcgca
      901 gttgagaagt ttcaacattt gcaataacaa gctttcaggt cctttgccag aagagattgg
      961 tgatttatac aatctagaag agcttgtggc ttatactaat aacttgaccg gtccattgcc
     1021 tcgttcttta ggaaatctca acaagttgac gacatttcga gcagggcaga acgatttctc
     1081 ggggaatata cctactgaaa ttggtaaatg tcttaacttg aagttgctcg gtcttgctca
     1141 gaatttcatt tctggagagc ttcccaagga aattgggatg cttgttaaac ttcaggaggt
     1201 gattctgtgg cagaacaagt tttcagggtt tattccgaaa gatattggga atttaacaag
     1261 tcttgagact cttgctttgt acggtaactc attggttgga cctataccat cagagattgg
     1321 taacatgaag tccttgaaga agctctatct ttaccaaaac cagttgaatg ggactatccc
     1381 aaaggagctt ggaaagcttt ccaaagtaat ggaaatcgat ttctcggaga atttgttgag
     1441 tggggagatt ccagttgagc tgagcaagat cagtgagttg aggctgctct atttgtttca
     1501 gaacaagtta actggaatca taccaaatga gctcagtaag ttgaggaatt tggcgaaact
     1561 cgatctctcc atcaactcct taaccggtcc tattccaccg ggtttccaga atcttacgtc
     1621 tatgcgccaa cttcagcttt tccacaattc cctcagtggt gtaatccctc aaggtcttgg
     1681 tttgtacagt cctctatggg ttgttgactt ctcagaaaac caactttcag gaaaaatacc
     1741 gccttttatc tgtcaacagt ctaatctcat tctgttgaat ctcgggtcta acaggatatt
     1801 tggaaatatt ccaccaggtg tattgcgctg caaatctctg ctgcagcttc gtgttgttgg
     1861 aaaccgactc actggtcagt ttccgactga gttatgcaag cttgtcaacc tttctgctat
     1921 tgagttggat caaaaccggt ttagcggacc actacctcct gagataggca cgtgtcagaa
     1981 gctgcaaaga cttcatcttg cagctaacca attttcctct aatctaccca acgaaatcag
     2041 taaactctcc aacctggtaa catttaacgt ttcctcaaac tctctcactg gaccaatccc
     2101 ctctgaaata gccaactgta aaatgcttca acgacttgac ttgagccgga acagtttcat
     2161 cggttcttta cctcccgagc ttggaagcct tcatcagctc gagatactca gactctcaga
     2221 aaatagattc tctggcaaca taccgtttac cattggaaac cttacccatc taactgagct
     2281 gcagatggga ggaaacttgt tttcaggaag tataccgcct caactcggtt tactctcgag
     2341 tctgcagatt gcaatgaacc tgagctacaa tgatttcagt ggtgaaatac ctcccgagat
     2401 cgggaatctt catctgctaa tgtacctatc tctgaacaac aaccatctca gcggcgaaat
     2461 cccgaccaca ttcgagaact tatccagttt actcgggtgc aacttttcct acaacaacct
     2521 caccggacag cttccacata cgcagatctt tcaaaacatg actctaacca gttttctcgg
     2581 aaacaaagga ctctgtggcg gccatttgag aagctgtgac cctagccact cctcttggcc
     2641 tcacatttca tctctaaaag cgggttctct ggaggtgctg ttccagggac cagggagcga
     2701 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     2761 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     2821 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     2881 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2941 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     3001 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     3061 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     3121 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     3181 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     3241 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     3301 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     3361 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     3421 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     3481 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     3541 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3601 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3661 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3721 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3781 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3841 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5941 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6001 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6061 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6121 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6181 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6241 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6301 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6361 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
