LOCUS       sequence-274575-        6593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G01820 X036_pECIA14 should be used with bait vector 
            AT2G01820 X036_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6593)
  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 6593)
  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: AT2G01820 X036_pECIA14
FEATURES             Location/Qualifiers
     source          1..6593
                     /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             1889..1912
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3587..3610
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3626..3643
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3666..3683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3716..3850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3740..3759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3794..3813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3994..4010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3994..4010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4007..4029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4018..4034
                     /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(4042..4048,4049..4066,4067..4072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4087..4108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4225..4242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4396..4984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4476..4495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5155..5946,5947..6015))
                     /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     5778..5797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6016..6120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6188..6206
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6244..6266)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6366..6385
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6579..6593,1..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 tctcaaaccg gtctcgatga
      541 ttcgactatg caatctctca aatcaagtct gaatcttaca tcagacgtcg attggtctaa
      601 ccctaaccct tgtaaatggc aatctgttca atgcgatgga agtaatcgtg ttactaagat
      661 tcagcttaaa caaaaaggga ttcgtggaac tcttcctact aatcttcaga gtctctctga
      721 attggttatt ctcgagcttt ttctcaatcg tatctctggt cctattcctg atctctctgg
      781 cttatctcgt ttacagactc tgaatctaca cgacaatctt tttacctctg taccgaagaa
      841 tctcttctct ggtatgagtt ctcttcaaga aatgtatctc gagaacaatc cgtttgatcc
      901 ttgggtaatt ccagacaccg ttaaagaagc tacgtctctg cagaatctaa ctctctccaa
      961 ttgcagcatt attggaaaga tcccagattt cttcggttct cagtcacttc cgagcttgac
     1021 gaatctgaaa ctatctcaga atggtttaga aggagagtta ccgatgagtt ttgctggtac
     1081 ttcgattcag tctctgttct tgaatggtca gaagcttaat ggatcgatct cggtattggg
     1141 gaacatgact tcgctcgttg aggtttctct tcaaggaaat cagttctcag gtccaattcc
     1201 tgatctctct ggtttggtat ctctacgtgt gtttaatgtt agagagaatc agctcactgg
     1261 tgttgttcca cagtctttgg ttagtttgag ttcgcttact actgtgaatt taactaataa
     1321 ttatcttcaa ggaccgactc cgttgtttgg caaatccgtt ggtgttgata tagtcaacaa
     1381 catgaatagc ttttgtacta atgtagctgg tgaggcttgt gatcctcgtg tggatacttt
     1441 ggtatctgta gctgaatcat ttggatatcc agtgaagctt gcggaaagtt ggaaaggaaa
     1501 caatccttgt gttaactggg ttgggattac ttgttctgga ggtaacatta cggttgttaa
     1561 tatgaggaaa caggatctct cgggtacgat ttctccgagt ttagctaagc ttacttcatt
     1621 ggagactatc aatcttgctg ataataaact ctccgggcat atacctgatg agcttaccac
     1681 attgtctaag cttaggttac ttgatgtgtc taacaacgat ttttatggaa taccgccaaa
     1741 gtttcgggat actgtgactt tggtgactga agggaacgct aacatgggga agaatggacc
     1801 taacaaaacg agtgatgctc ctggagcttc accaggaagt aaaccttcag gaggaagtga
     1861 tggcagtgaa acttctaaga aatcaagcct ggaggtgctg ttccagggac caagcggcaa
     1921 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     1981 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     2041 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2101 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2161 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     2221 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     2281 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2341 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2401 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2461 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     2521 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     2581 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     2641 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     2701 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     2761 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     2821 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     2881 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     2941 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3001 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3061 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3121 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3181 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     3241 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3301 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3361 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3421 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3481 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     3541 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     3601 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3661 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3841 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3901 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     3961 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4021 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4081 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4141 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4201 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4261 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4981 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5041 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5101 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5161 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5221 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5281 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5341 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5401 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5461 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5521 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5581 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5641 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5701 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5761 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5821 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5881 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5941 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6001 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6061 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6121 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6181 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6241 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6301 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6361 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6421 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6481 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6541 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
