LOCUS       sequence-276515-        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G39270 K12_pECIA14 should be used with bait vector 
            AT4G39270 K12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  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 6380)
  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: AT4G39270 K12_pECIA14
FEATURES             Location/Qualifiers
     source          1..6380
                     /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             1676..1699
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3374..3397
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3413..3430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3453..3470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3503..3637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3527..3546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3581..3600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3781..3797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3781..3797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3794..3816)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3805..3821
                     /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(3829..3835,3836..3853,3854..3859))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3874..3895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4012..4029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4183..4771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4263..4282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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     5565..5584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5803..5907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5975..5993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6031..6053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6153..6172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6366..6380,1..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 agttctctga atgagaaatc
      541 ggcgcttctg ctgttaaggt cttctcttgg gttacgaagc agagattggc ctgtaaaagg
      601 caatccttgt ctgaactgga acggcatcaa atgtgatcag aatggtagag taactaagat
      661 caacatttca gggttcagaa ggacaaggat tgggaatcaa aacccggaat tctcagttgg
      721 ttctctcgtc aatctcaccc gtctagcttc cttcaacgcc tcaaggttct atcttccagg
      781 tccaatcccg gctttgtttg gatcgagtct gttgacttta gaggtgttgg accttagctc
      841 ctgttcaatc acaggaacca tccctgaaag tttgactcgg ttatcacatc tcaaagttct
      901 tgatctttca aagaatgcga tcaatgggga tattcctctg tctcttacct ctcttcagaa
      961 tctatcaatc cttgatcttt cctcaaactc ggtgtttggt tcgattcctg ctaatattgg
     1021 ggcactctca aagctccagc gtctgaatct ttcgcgtaac accttgactt cttcaatacc
     1081 tccatcactc ggagacctct ctgttttaat tgaccttgat ctcagcttca acggaatgtc
     1141 aggttcggtt ccatcagatt tgaaagggct aaggaacttg cagacattgg taattgcggg
     1201 aaatcgcctt tcaggatccc tacctcctga tctgtttagt ttgctgagta aacttcagat
     1261 cattgacttc agaggcagtg gtttcatagg cgctttaccg tccaggttat ggtcgttacc
     1321 ggagctcaag tttctagata tttctggaaa tcacttctct gacatgctgc ctaataccac
     1381 tgtcagtttt gattctactg tctccatgct taacatatcc gggaacatgt tttacggcaa
     1441 tctcacactt ttactgacaa ggttccaagt tgttgacctg tcagaaaact attttgaagg
     1501 taaaattcct gactttgtgc caaccagagc ttcattgagc aacaattgtc ttcagggccc
     1561 agagaagcag cggaagttgt cggattgcac tttgttttat tcaaaaaagg gcttaacttt
     1621 caacaatttc ggacaacatg aagagaagaa gagttctaaa acttcctggt taagcctgga
     1681 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1741 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1801 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1861 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1921 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1981 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     2041 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     2101 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     2161 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     2221 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     2281 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     2341 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     2401 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     2461 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2521 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2581 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2641 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2701 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2761 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2821 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2881 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2941 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     3001 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     3061 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     3121 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     3181 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     3241 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     3301 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     3361 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     3421 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3481 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3541 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3601 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3661 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3721 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3781 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3841 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3901 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4621 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5221 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5941 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6001 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6061 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6361 gttttcccag tcacgacgtt
//
