LOCUS       sequence-276504-        7373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G33170 X049_pECIA14 should be used with bait vector 
            AT2G33170 X049_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  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 7373)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..7373
                     /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             4367..4390
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4406..4423
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4446..4463)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4496..4630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4520..4539)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4574..4593
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4774..4790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4774..4790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4787..4809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4798..4814
                     /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(4822..4828,4829..4846,4847..4852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4867..4888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5005..5022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5256..5275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5935..6726,6727..6795))
                     /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     6558..6577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6796..6900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6968..6986
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7024..7046)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7146..7165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7359..7373,1..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 caagcggcaa
     2701 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     2761 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     2821 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2881 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2941 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     3001 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     3061 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     3121 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     3181 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     3241 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     3301 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     3361 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     3421 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     3481 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     3541 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     3601 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     3661 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     3721 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3781 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3841 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3901 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3961 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     4021 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     4081 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     4141 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     4201 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     4261 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     4321 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     4381 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     4441 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     4501 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4561 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4621 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4681 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     4741 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4801 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4861 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4921 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6961 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     7021 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7081 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7141 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7201 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     7261 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7321 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
