LOCUS       sequence-298978-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G16250 K9_pECIA2 should be used with prey vector 
            AT2G16250 K9_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  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 5504)
  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: AT2G16250 K9_pECIA2
FEATURES             Location/Qualifiers
     source          1..5504
                     /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             1757..1780
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2486..2527
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2537..2554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2577..2594)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2627..2761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2651..2670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2705..2724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2905..2921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2905..2921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2918..2940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2929..2945
                     /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(2953..2959,2960..2977,2978..2983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3136..3153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3307..3895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3387..3406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4066..4857,4858..4926))
                     /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     4689..4708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4927..5031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5099..5117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5155..5177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5277..5296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5490..5504,1..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 agtcgattca gtgagaaact
      541 cattttgctt aatctcaggt cttctttagg gttaagagga acagattggc caattaaagg
      601 tgacccttgt gttgattgga gaggtataca atgtgagaat ggtagtatca ttgggatcaa
      661 tatctctggg tttaggagaa caagaattgg taagctaaac ccacaattct ctgttgatcc
      721 tcttcgtaat ctaactcgtt tgtcgtattt taacgcgtcc gggttagcgt taccggggac
      781 tatacctgag tggtttggtg ttagtttgtt ggcattggaa gtattggatc ttagttcgtg
      841 ttctgttaac ggtgttgttc cttttactct tggtaatctc actagtttaa gaactttgaa
      901 tctgtctcag aacagtctta ctagtttagt tcctagcagt ttaggacagt tgttgaatct
      961 atcacagctt gatctctcta ggaactcatt cactggtgtt cttcctcaat cttttagctc
     1021 tctgaagaat ctgttgactc ttgatgtttc ttccaattac cttactggac ctatccctcc
     1081 tggtttagga gcgttatcaa agcttatcca tctgaacttt tcaagcaata gtttctcgtc
     1141 tccgattcct tcagagcttg gtgatcttgt caatctggtt gactttgatc tcagcatcaa
     1201 ttcattgtct ggttcagttc ctcaggagct aagaaagtta agtaagttgc agctaatggc
     1261 gattggcgat aaccttttga gtggtacttt accagtagat ttgtttagtg ctgagagcca
     1321 actgcaaact ttagttttga gggaaaatgg tttctctggt agtctacctg atgtgtgctg
     1381 gtcgttgcca aaattgcgga ttcttgacat tgctaagaac aacttcacag gactgctgcc
     1441 atattctagt tatgattcag accagatagc tgaaatggtt gatatctctt cgaacacatt
     1501 ctatggagag ctcacaccga ttcttagacg gttcaggata atggacctct ctggtaatta
     1561 tttcgaaggt aagcttcctg attatgtgac cggggaaaat gtgtctgtta caagcaattg
     1621 ccttcgaaat gagaggagac agaagccgtc agcaatatgt gcagcattct ataagtctag
     1681 aggattagat tttgatgatt tcggacgtcc taatttgaca caaccgacat cgaaaaatgc
     1741 ttcttcaggg ataagcctgg aggtgctgtt ccagggacca gggagcgaca aaactcacac
     1801 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     1861 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     1921 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     1981 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     2041 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     2101 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     2161 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     2221 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     2281 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     2341 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     2401 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     2461 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     2521 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     2581 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     2641 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2701 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2761 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2821 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     2881 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3061 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3481 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3721 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5041 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5101 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5161 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5221 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5281 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5341 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     5401 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     5461 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
