LOCUS       sequence-275916-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G74360 X038_pECIA14 should be used with bait vector 
            AT1G74360 X038_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  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 7301)
  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: AT1G74360 X038_pECIA14
FEATURES             Location/Qualifiers
     source          1..7301
                     /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             2597..2620
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4295..4318
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4334..4351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4374..4391)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4424..4558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4448..4467)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4502..4521
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4702..4718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4702..4718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4715..4737)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4726..4742
                     /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(4750..4756,4757..4774,4775..4780))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4795..4816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4933..4950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5104..5692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5184..5203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5863..6654,6655..6723))
                     /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     6486..6505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6724..6828)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6896..6914
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6952..6974)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7074..7093
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7287..7301,1..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 tctctggaca gtgatagaga
      541 agtcttgtta agcctaaaat cgtaccttga atcacggaac ccacagaacc gaggactata
      601 cacggaatgg aaaatggaga accaagatgt tgtgtgtcaa tggcctggaa tcatatgtac
      661 acctcagaga agtagagtta cagggatcaa tctaactgat tccaccattt caggtcctct
      721 attcaagaac ttctccgctt taacggagct cacatatctc gatttatcaa ggaatacgat
      781 cgaaggggag attccagatg acttgagccg ttgtcacaac ttaaagcatc tgaatctttc
      841 tcataatatc cttgaaggag agctaagttt acccgggttg tcaaatcttg aggttcttga
      901 tctttcgttg aataggatta cgggtgatat tcagtctagc ttcccattgt tctgcaacag
      961 tttggttgtt gcaaatctgt cgaccaacaa ctttactgga agaatcgatg acatcttcaa
     1021 cgggtgtagg aatctcaagt atgttgactt cagttccaat agattcagtg gagaagtgtg
     1081 gactggtttt gggaggctgg tcgagttttc agttgctgat aatcatctct ccgggaatat
     1141 ctctgcttca atgttcaggg ggaactgtac tttgcaaatg ttggatttgt ctgggaacgc
     1201 ttttggtggg gaatttccag gccaagtttc gaattgccag aatctaaatg tactgaatct
     1261 ttggggaaac aagttcacag ggaacattcc agctgagatt ggttccatat cctctctcaa
     1321 aggtttgtac ttggggaata atacattttc tcgagacata ccggaaactc tcctgaactt
     1381 gaccaacttg gtgtttctgg atttgagcag aaacaaattt ggaggagaca ttcaagaaat
     1441 ctttgggaga ttcacgcaag taaagtatct agtcttgcat gcaaactcat acgtaggagg
     1501 catcaattct tccaacatcc tcaagttacc caatctttca aggctagatt tgggctacaa
     1561 caatttctcg ggacagttac ctactgaaat ttctcagata cagagtttga agttcttgat
     1621 tcttgcttat aataacttca gtggcgatat accacaggag tatgggaaca tgccggggct
     1681 tcaagcactt gatctctcct ttaacaagct gaccggttcg ataccagctt catttgggaa
     1741 attgacctct cttttgtggc taatgcttgc aaacaactct ctatcaggag aaatccctcg
     1801 agagattggc aactgcacaa gccttttgtg gtttaacgtg gcaaacaacc agctctctgg
     1861 tagattccat cctgaattga ctagaatggg gagtaatcct tctccaacat ttgaagtgaa
     1921 taggcaaaac aaagacaaga taattgctgg ttctggtgaa tgcttggcga tgaagagatg
     1981 gattccagcg gagttccctc cgttcaactt tgtatacgca attctaacca aaaagagttg
     2041 cagaagccta tgggaccatg ttctcaaagg gtacggtctc tttcctgtct gttctgctgg
     2101 ttccacggta cgcacgctta aaatttcagc ctatcttcag ctcagtggaa ataagttttc
     2161 aggtgagatt cctgcaagca tctctcagat ggacaggctg agcacactac atttgggttt
     2221 caatgagttt gaggggaaac tgcctccaga gataggacaa ttgcctcttg cattcctcaa
     2281 ccttactcga aacaatttct cgggcgagat tcctcaagaa atcggaaatc tgaagtgcct
     2341 gcagaatctt gatctgtctt tcaacaattt ctctggaaac tttccaacga gtttgaatga
     2401 cttgaatgag ctgagtaagt tcaacatctc atataaccct ttcatttctg gcgcgatacc
     2461 aacaacagga caagtagcaa cctttgacaa agactccttt cttggaaatc cgctgctgcg
     2521 gtttcctagt ttcttcaacc aatcagggaa caacacgagg aagatttcca accaagtcct
     2581 aggaaacagg ccaagactgg aggtgctgtt ccagggacca agcggcaatc ccggcggcga
     2641 tctggccccc cagatgctgc gcgagctgca ggagaccaat gccgccctgc aggacgtgcg
     2701 cgagctgctg cgccaccgcg tgaaggagat caccttcctg aagaacaccg tgatggagtg
     2761 cgacgcctgc ggcatgcagc cctctagagg ctccggctcc atcatcccag ttgaggagga
     2821 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     2881 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     2941 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     3001 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     3061 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     3121 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     3181 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     3241 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     3301 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     3361 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
     3421 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     3481 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     3541 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     3601 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     3661 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     3721 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     3781 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     3841 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     3901 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     3961 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     4021 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     4081 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     4141 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     4201 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     4261 cccgggttct agaggaagca cctccggcga taacgattac aaggatgatg atgataaggc
     4321 cagcggcacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     4381 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     4441 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4501 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4561 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4621 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga cctgcaggca
     4681 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4741 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4801 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5941 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6001 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6061 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6121 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6181 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6241 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6301 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6361 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6421 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6481 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6541 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6601 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6661 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6721 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6781 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6841 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6901 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6961 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7021 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7081 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7141 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7201 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7261 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
