LOCUS       sequence-274524-        6962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G53430 X031_pECIA14 should be used with bait vector 
            AT1G53430 X031_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6962)
  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 6962)
  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: AT1G53430 X031_pECIA14
FEATURES             Location/Qualifiers
     source          1..6962
                     /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             2258..2281
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3956..3979
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3995..4012
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4035..4052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4085..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4163..4182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4363..4379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4363..4379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4376..4398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4387..4403
                     /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(4411..4417,4418..4435,4436..4441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4456..4477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4594..4611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5524..6315,6316..6384))
                     /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     6147..6166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6385..6489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6557..6575
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6613..6635)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6735..6754
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6948..6962,1..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 caacttttgc ctgaagatga
      541 agttcaaaca ttgcgaacga tctttagaaa gcttcaaaac caaacagtga acatcgaaag
      601 aacttcttgt tcggaccaaa actggaactt tgttgtcgag tcagcctcca actcaccaac
      661 cagtaacatt acctgcgact gtactttcaa cgccagctca gtctgtcgtg tcacaaacat
      721 acagcttaaa agtttcagct tgcccggaat attcccgcct gagttcggaa acctcacgcg
      781 tcttcgagag atagatcttt cgcggaactt tctcaatgga acaataccta cgacattgtc
      841 tcaaattcca cttgaaatct tgtctgtaat cggaaaccgt ctctcaggac catttcctcc
      901 tcaacttgga gatattacta cacttactga tgtgaatttg gaaactaatt tattcacagg
      961 accacttcct cgaaacctag ggaacttaag aagcttgaaa gagttacttc tctctgcaaa
     1021 caacttcacg ggtcaaatcc ctgagtcctt aagtaatctc aagaatttga ctgaatttcg
     1081 gattgatgga aactctctat ctgggaaaat acctgatttt attggaaact ggactctgct
     1141 ggagaggcta gaccttcaag gcacatcaat ggaaggtcca attccacctt caatatcaaa
     1201 cttgacgaac ttgactgaat tgaggataac cgatttgcgt ggacaagcgg ccttttcttt
     1261 tccggacctg cgaaatctaa tgaaaatgaa acgattggta ctaagaaatt gtttgataag
     1321 gggacctatc ccggaataca ttggttccat gagtgagctg aagacactag atttaagctc
     1381 aaacatgcta accggtgtaa ttcctgacac atttcggaat ctcgatgcat ttaactttat
     1441 gtttttgaat aataactcat tgactggtcc agttcctcag ttcatcatca acagtaaaga
     1501 aaacctagat ttatctgaca acaatttcac tcagccacct actttaagct gtaatcagct
     1561 tgatgtgaac ttgatctcca gctacccttc agtaaccgat aactctgttc aatggtgctt
     1621 gagagagggt cttccatgtc cagaagatgc taaacagtct tccctgttca tcaactgtgg
     1681 aggaagcaga ctcaagattg gaaaagacac ttatacagat gacttgaaca gtagaggaca
     1741 atcaacattc tcttctgtct ctgaaagatg gggatacagt agttctggag tttggttagg
     1801 caaggaggat gctggctact tagcaacaga cagatttaac ttgatcaatg gatcaactcc
     1861 agagtattac aaaacagccc gtctctctcc acagtcactc aagtactatg gactatgctt
     1921 gagaagagga agttacaaac tgcagctaca ttttgcagag ataatgttct caaatgacca
     1981 gacttttaat agcttagggc ggcgaatatt cgacatttat gtccaaggga acttgttgga
     2041 gagggacttt aacatagcag agagagcagg tggagttggt aaaccgttca taaggcaaat
     2101 tgatggagtt caagtgaatg gaagtacgtt agagattcat ttgcagtgga cagggaaagg
     2161 cacaaacgta ataccaacaa gaggtgttta cgggcctctc atatctgcca taaccattac
     2221 accaaatttc aaggttgata ccggaaaacc attgtccctg gaggtgctgt tccagggacc
     2281 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     2341 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     2401 gaagaacacc gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc
     2461 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     2521 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     2581 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     2641 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     2701 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     2761 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     2821 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     2881 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     2941 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     3001 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     3061 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     3121 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     3181 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     3241 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     3301 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     3361 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     3421 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     3481 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     3541 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     3601 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     3661 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     3721 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     3781 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     3841 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     3901 cggcaccacc gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta
     3961 caaggatgat gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta
     4021 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     4081 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4141 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4201 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4261 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     4321 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4381 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4441 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4501 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4561 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4621 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5581 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5641 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5701 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5761 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5821 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5881 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5941 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6001 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6061 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6121 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6181 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6241 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6301 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6361 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6541 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6601 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6661 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6721 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6781 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6841 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6901 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6961 tt
//
