LOCUS       sequence-270411-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G23300 X009_pECIA14 should be used with bait vector 
            AT2G23300 X009_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  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 6122)
  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: AT2G23300 X009_pECIA14
FEATURES             Location/Qualifiers
     source          1..6122
                     /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             1418..1441
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3116..3139
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3155..3172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3195..3212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3245..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3323..3342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3523..3539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3523..3539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3536..3558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3547..3563
                     /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(3571..3577,3578..3595,3596..3601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3616..3637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3754..3771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3925..4513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4005..4024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4684..5475,5476..5544))
                     /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     5307..5326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5545..5649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5717..5735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5773..5795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5895..5914
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6108..6122,1..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 aactctgatg gagttctctt
      541 actgagtttc aaatactctg ttcttcttga tcctctctct ttattacaat catggaacta
      601 cgaccacgac aatccttgtt catggcgagg tgtgttgtgt aataacgatt caagagttgt
      661 tactttatct ctcccaaact ctaacctcgt tggttcgatt ccttccgatc tgggtttcct
      721 ccaaaacctc caaagtctta atctttccaa taattcactc aatgggtcat taccggttga
      781 gtttttcgcc gccgataagc tccggtttct tgatttatca aataacttga tctccggcga
      841 gatccctgta tcaatcggag gtttacacaa cctccagacg ttaaatctct ccgataacat
      901 cttcaccggg aaactaccag ctaacttagc gtctcttgga agcttaacgg aggtttctct
      961 gaagaacaac tacttctccg gcgagtttcc cggcggcgga tggagatcgg ttcagtatct
     1021 agacatttct tcaaatctaa tcaacggttc actcccacct gatttctccg gcgacaatct
     1081 ccgatacctg aatgtctcgt ataaccaaat ctccggagag attcctccga atgttggtgc
     1141 cggttttcct caaaacgcca ccgttgattt ctccttcaac aatttaaccg gttcaatccc
     1201 agattctccg gtttacctta accagaaatc aatttcgttt tccggaaacc cgggtttatg
     1261 cggaggtccg acccgaaacc cgtgtcccat tccttcatct ccggccaccg tctcgccacc
     1321 aacctctaca cctgcactcg cagctatacc taaatcaatc gggtctaatc gagaaaccga
     1381 accgaacaac aactcaaatc ctcgaaccgg gttaagactg gaggtgctgt tccagggacc
     1441 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     1501 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     1561 gaagaacacc gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc
     1621 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     1681 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     1741 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     1801 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     1861 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     1921 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     1981 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     2041 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     2101 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     2161 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     2221 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     2281 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     2341 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     2401 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     2461 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     2521 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     2581 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     2641 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     2701 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     2761 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     2821 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     2881 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     2941 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     3001 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     3061 cggcaccacc gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta
     3121 caaggatgat gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta
     3181 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     3241 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3301 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3361 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3421 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     3481 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     3541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3781 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4501 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4621 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5701 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5761 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5821 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5881 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5941 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6001 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6061 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6121 tt
//
