LOCUS       sequence-296640-        5789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT4G20450 N11_pECIA2 should be used with prey vector 
            AT4G20450 N11_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5789)
  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 5789)
  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: AT4G20450 N11_pECIA2
FEATURES             Location/Qualifiers
     source          1..5789
                     /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             2042..2065
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2771..2812
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2822..2839
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2862..2879)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2912..3046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2936..2955)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2990..3009
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3190..3206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3190..3206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3203..3225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3214..3230
                     /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(3238..3244,3245..3262,3263..3268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3283..3304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3421..3438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3592..4180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3672..3691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4351..5142,5143..5211))
                     /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     4974..4993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5212..5316)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5384..5402
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5440..5462)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5562..5581
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5775..5789,1..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 caaggattca tcagcttaga
      541 ttgcggaatg ccccgtaacg agtctagtta taccgacgag tccaccggtt tgaatttctc
      601 atccgacgct gattttatta gcagcggaaa aagtggtacc atcaagaccg aagactctga
      661 ttctggcgtg aaatacataa agccatataa gcagcttagg tattttcctg aaggagcacg
      721 aaactgctat aatctgactg tcatgcaggg cacacattat cttatcagag ccgtgttcgt
      781 atatggaaat tatgacctca aacaaagacc aaaatttgat ctatatcttg gccccaattt
      841 ctggacaact ataaatttgc aagatccttc gggtggtttc tattatcgga tttggttgca
      901 agatggtacc gtagaggagg taatccacat gccaaagtca aacaatttgg atatttgttt
      961 ggttaagaca ggaacaacga cgccattcat atcatccttg gaactacgtc ccttgagaga
     1021 tgacacctat actactacaa ccggctctct aaagctaatc tcccgctggt atttcaggaa
     1081 gcctttcccc actcttgaat ccatcatacg acatccagat gatgtccatg accgtctatg
     1141 ggacgtatac catgcggatg aagaatggac agatataaac accaccaccc ctgtaaatac
     1201 aactgttaat gcctttgatc tgccacaagc tataatttca aaggcttcca taccccaagt
     1261 tgcaagtgat acatggagca caacctggtc gatacaaaac ccggacgacg acgttcacgt
     1321 gtaccttcac tttgccgaga tccaagcctt gaagcccagt gataccagag agtttagtat
     1381 cttgtggaat aaaaacacaa tcattaggga ttactacagt cctctagagt tcatggctga
     1441 cactgtacct ataagaacct cgagtaaatg cggtgacgat ggtttctgct ccctagatct
     1501 cacaagaact aagtcgtcaa ctcttccacc atattgtaat gcgatggagg ttttcgggtt
     1561 actccagctt cttcagaccg aaacagatga aaacgacgtc actacattga agaacattca
     1621 agccacttat agaatacaaa aaacgaactg gcaaggagat ccatgtgtac ctatacagtt
     1681 tatatggact ggcctaaact gtagcaacat gtttccgtct attccaccaa gaatcacttc
     1741 catagacttt tctaactttg gattaaatgg gaccataaca agtgatatac aatatcttaa
     1801 ccagctacag aaattggacc tgtcaaataa caacttgacg ggaaaggtgc ctgaatttct
     1861 cgccaaaatg aagttgttga cattcataaa cttgagtggg aacaatctta gcggctcaat
     1921 tcctcaaagc cttctcaaca tggaaaagaa tgggctaata acgttgctgt ataatggaaa
     1981 taacctctgt ttggatccat catgcgaaag tgaaactggt cctgggaata acaaaaagaa
     2041 actggaggtg ctgttccagg gaccagggag cgacaaaact cacacatgcc caccgtgccc
     2101 agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac
     2161 cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga
     2221 ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa
     2281 gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca
     2341 ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc
     2401 ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac
     2461 cctgccccca tcccgggagg agatgaccaa gaaccaggtc agcctgacct gcctggtcaa
     2521 aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa
     2581 ctacaagacc acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct
     2641 caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga
     2701 gggtctgcac aaccactaca cgcagaagag cctctccctg tctccgggta aatctagagg
     2761 gcccttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2821 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     2881 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     2941 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3001 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3061 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3121 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     3181 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3241 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3301 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3361 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3421 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3481 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3541 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3781 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3961 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4021 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4081 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4261 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4321 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4441 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4501 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4561 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4861 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5341 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5401 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5461 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5521 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5581 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5641 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5701 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5761 aacgccaggg ttttcccagt cacgacgtt
//
