LOCUS       sequence-275976-        7349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G17230 X074_pECIA14 should be used with bait vector 
            AT1G17230 X074_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7349)
  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 7349)
  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: AT1G17230 X074_pECIA14
FEATURES             Location/Qualifiers
     source          1..7349
                     /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             2645..2668
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4343..4366
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4382..4399
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4422..4439)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4472..4606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4496..4515)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4550..4569
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4750..4766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4750..4766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4763..4785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4774..4790
                     /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(4798..4804,4805..4822,4823..4828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4843..4864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4981..4998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5152..5740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5232..5251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5911..6702,6703..6771))
                     /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     6534..6553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6772..6876)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6944..6962
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7000..7022)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7122..7141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7335..7349,1..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 tccttaaacg aagaagggcg
      541 tgttcttctg gagtttaaag ctttccttaa tgactccaat ggctatctcg caagctggaa
      601 tcagttggat tcaaatccct gcaactggac cggaatcgcg tgtacccatc tcagaactgt
      661 aacttctgtt gatctaaatg ggatgaatct atcaggtaca ctatctcctc tcatatgcaa
      721 gcttcatggt ttaaggaaat tgaatgtctc caccaacttc atctctggtc caatccctca
      781 agatttatct ctctgtcgaa gtctagaggt tcttgatctc tgcacaaata ggttccatgg
      841 agtgatacca attcagctaa ccatgatcat cactcttaag aagctgtatc tatgcgagaa
      901 ttatcttttc ggttctatcc ctagacaaat cggtaatttg agttcactcc aagagcttgt
      961 gatctacagc aataatctca ccggtgtaat ccctccttcc atggcgaaat tgagacagct
     1021 ccggatcatt agggcaggac ggaatggatt ctcgggtgtg attccgtctg aaatcagcgg
     1081 atgcgagagc ctaaaggtac ttggattagc agagaatctt cttgagggtt ctcttcctaa
     1141 gcagctcgag aagcttcaaa atcttactga tttgatactc tggcaaaacc gtctatctgg
     1201 tgagatacct ccttccgtag gaaacatcag cagactggag gtgcttgcgc tgcatgaaaa
     1261 ctacttcacg ggatcaattc caagagagat tgggaagctc acaaagatga aaaggctgta
     1321 cctttacaca aaccagttaa ccggagagat acctcgcgaa ataggcaact tgatagatgc
     1381 agctgagata gatttttcgg agaatcagtt aacaggtttc ataccaaaag agtttggtca
     1441 tatcctgaat ctcaaattac ttcatctttt tgagaacatt cttctaggtc caattcctag
     1501 ggagcttggg gaattgacac tgttggagaa gctagacttg tctatcaata gactaaacgg
     1561 tactatcccg caagagcttc agttcctacc ttaccttgta gatttgcagc tttttgacaa
     1621 ccagcttgaa gggaagatac ctcctttaat cggtttctat agcaattttt ctgttcttga
     1681 catgtctgcc aattctctgt ctggtccaat tcctgcccac ttctgcagat ttcagacgct
     1741 aatacttctg agtcttggtt caaacaagtt atcaggaaac atccctcgtg atctgaaaac
     1801 ctgcaagtca ctgacaaagc taatgctagg cgataaccag ctaacaggaa gccttcccat
     1861 tgaattattt aatcttcaga accttactgc attagagctt caccaaaact ggttatcagg
     1921 gaatatatct gcagacctgg gaaagctgaa gaatttggaa agactgcgtc tagctaacaa
     1981 caattttact ggtgaaattc ctcctgagat tgggaatctc acaaagattg ttggctttaa
     2041 catctcctcc aatcagctca ctggtcacat ccctaaagag ttgggaagtt gtgtcactat
     2101 ccagaggctt gaccttagtg gcaacaagtt ttccggatac atagcacaag agcttggtca
     2161 gttggtatat ttggagattt tgagattgtc tgataataga ttgaccggag aaattcctca
     2221 tagctttgga gatcttactc ggctcatgga gttacaacta ggaggcaatc tcttgtctga
     2281 aaacatccca gtggagctcg ggaagttaac ctctctgcaa atctccctca acattagtca
     2341 caataatctc tcaggtacta ttccagacag tttaggaaat ctacagatgt tggagatact
     2401 ttacctcaac gacaacaagc tttcaggtga gatccctgca tcaattggga atctcatgag
     2461 cctcctcatc tgcaatatct caaacaataa cttggtcgga acagtaccag acacagccgt
     2521 gttccagcgg atggattcat cgaactttgc tggaaaccat ggtttgtgta actctcagag
     2581 aagccattgt caaccacttg tcccacattc tgattcaaag ctaaattggt taatcaacgg
     2641 ttcgctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     2701 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     2761 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     2821 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     2881 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     2941 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     3001 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     3061 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     3121 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     3181 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     3241 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     3301 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     3361 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     3421 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     3481 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     3541 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     3601 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     3661 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     3721 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     3781 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     3841 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     3901 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     3961 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     4021 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     4081 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     4141 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     4201 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     4261 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     4321 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     4381 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     4441 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     4501 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4561 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4621 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4681 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     4741 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4801 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4861 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4921 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4981 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5041 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5821 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5941 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6001 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6061 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6121 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6181 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6241 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6301 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6361 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6421 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6481 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6541 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6601 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6661 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6721 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6781 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6901 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6961 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7021 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7081 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7141 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7201 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7261 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7321 aacgccaggg ttttcccagt cacgacgtt
//
