LOCUS       sequence-270522-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G22730 K11_pECIA14 should be used with bait vector 
            AT4G22730 K11_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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: AT4G22730 K11_pECIA14
FEATURES             Location/Qualifiers
     source          1..6080
                     /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             1376..1399
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3074..3097
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3113..3130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3153..3170)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3203..3337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3227..3246)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3281..3300
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3481..3497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3481..3497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3494..3516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3505..3521
                     /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(3529..3535,3536..3553,3554..3559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3574..3595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3712..3729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4642..5433,5434..5502))
                     /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     5265..5284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5503..5607)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5675..5693
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5731..5753)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5853..5872
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6066..6080,1..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 cgaggaaatg cagagctgaa
      541 ggctttaatg gagctgaaat cgtcgcttga ccctgaaaac aagcttctcc gttcatggac
      601 gtttaacggc gatccatgcg acggatcttt cgaaggaatt gcatgtaacc agcatctaaa
      661 agtcgcaaac atatcattac aagggaaacg tttggtcgga aaattgtctc cggcggttgc
      721 agagctcaaa tgtttgtcag gtctttactt acattacaat agtctctctg gagagatacc
      781 tcaagagatc acaaatctta ctgaattatc agatctttat ctcaatgtta ataacttctc
      841 tggtgagatt ccggcagata tcggctccat ggctggcttg caagttatgg atctttgttg
      901 caacagttta acagggaaga taccaaagaa cattggatcc ttgaagaaac ttaatgtgtt
      961 gtctctgcaa cacaacaaac taaccggaga ggttccttgg actttaggaa acttgagtat
     1021 gttaagcagg cttgatttaa gcttcaacaa tctgttaggt ttaatcccaa aaaccctagc
     1081 caacattcct cagttggaca ctcttgactt gcgcaacaat actctctctg gctttgttcc
     1141 tcctggtctt aagaagttga atgggagctt ccagtttgag aacaacaccg ggttatgtgg
     1201 tatcgatttt ccttctttga gagcttgttc tgctttcgat aatgcgaata atatcgaaca
     1261 atttaagcag cctcctggtg aaatagacac tgataaatca gctcttcaca acattcccga
     1321 gtctgtatat ctccaaaagc attgcaacca aacacattgc aagaaatcct catcgctgga
     1381 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1441 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1501 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1561 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1621 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1681 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     1741 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     1801 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     1861 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     1921 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     1981 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     2041 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     2101 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     2161 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2221 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2281 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2341 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2401 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2461 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2521 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2581 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2641 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     2701 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     2761 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     2821 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     2881 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     2941 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     3001 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     3061 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     3121 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3181 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3241 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3301 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3361 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3421 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5641 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5701 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5761 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5821 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5881 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6001 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6061 gttttcccag tcacgacgtt
//
