LOCUS       sequence-270158-        6665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G46420 N10_pECIA14 should be used with bait vector 
            AT3G46420 N10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6665)
  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 6665)
  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: AT3G46420 N10_pECIA14
FEATURES             Location/Qualifiers
     source          1..6665
                     /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             1961..1984
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3659..3682
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3698..3715
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3738..3755)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3788..3922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3812..3831)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3866..3885
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4066..4082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4066..4082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4079..4101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4090..4106
                     /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(4114..4120,4121..4138,4139..4144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4159..4180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4297..4314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4468..5056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4548..4567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5227..6018,6019..6087))
                     /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     5850..5869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6088..6192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6260..6278
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6316..6338)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6438..6457
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6651..6665,1..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 ggatttatca gtgtagactg
      541 cggattatct cctaatgagg tctctcctta tattgagccg tttactggac tacagttcac
      601 aacggattca aatttcattg aaaccggaaa acttggtaga attcaggcaa gcctagagcc
      661 gaagtaccga aagtcgcaaa cgacgttgag atattttcct gatggaatac gcaactgtta
      721 caatcttaca gtgacacaag gaaccaacta tttaatcaga gctagagcaa tatatgggaa
      781 ttatgatggt cttaacattt atcctaaatt tgacttgtac ataggcccta atttttgggt
      841 aacaatagac ttaggaaagt atgtaaatgg tacatgggag gagatcatct acataccaaa
      901 atctaatatg ttggacgtgt gtcttgtcaa gacagggccg tccacaccat tgatatcatc
      961 cttggtgtta aggcctcttg caaatgctac ttacatcact caatctggtt ggttgaagac
     1021 ttatgttcgg gtttatctaa gcgactcaaa tgacgtcata cgatatccgg atgatgtcta
     1081 tgatcgtata tggggttcat actttgagcc ggagtggaag aagatttcca ctacacttgg
     1141 agtgaacagt tctagtggat ttctgcctcc actaaaggca ctcatgactg ccgcctctcc
     1201 tgccaatgct agtgcaccat tggctattcc cggagtacta gatttcccaa gtgataaact
     1261 atacttgttc ctccacttct ctgagattca agtcttaaag gccaacgaaa ctagagagtt
     1321 cgaaattttt tggaataaaa agcttgttta caatgcttat agtcctgtat atctacagac
     1381 caagactata cgcaacccat ctccagttac ttgtgaaaga ggagaatgca tactagagat
     1441 gatcaaaact gagaggtcaa ctctcccgcc tttgcttaac gccgttgaag ttttcacagt
     1501 ggttgaattt cctcagccgg aaacagatgc tagtgatgtg gtggctataa aaaacatcaa
     1561 ggccatctat ggattgactc gagtcacatg gcaaggagat ccatgtgttc ctcaacaatt
     1621 tttgtggaac ggtttaaatt gcaacagcat ggagacgtct acacctccaa gaatcacttc
     1681 cttggacttg tcttcaagcg ggttaaccgg aagcatatca gttgttattc aaaatcttac
     1741 ccatctagaa aagttggatt tgtcaaataa caacttgact ggggaagttc ccgattttct
     1801 agcaaacatg aaatttcttg tgttcataaa cttaagcaag aacaatctga atggttcgat
     1861 tcctaaagct cttcgcgata gagaaaataa aggacttaag ttaattgtcg acaaaaatgt
     1921 ggacaactgc tcatctggtt catgcaccca aaaaaagaaa ctggaggtgc tgttccaggg
     1981 accaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     2041 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     2101 cctgaagaac accgtgatgg agtgcgacgc ctgcggcatg cagccctcta gaggctccgg
     2161 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     2221 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     2281 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     2341 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     2401 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     2461 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     2521 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     2581 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     2641 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     2701 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     2761 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     2821 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     2881 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     2941 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     3001 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     3061 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     3121 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     3181 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     3241 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     3301 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     3361 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     3421 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     3481 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     3541 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     3601 cgccggcacc accgacgccg cgcacccggg ttctagagga agcacctccg gcgataacga
     3661 ttacaaggat gatgatgata aggccagcgg caccggtcat catcaccatc accattgagt
     3721 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     3781 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3841 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3901 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3961 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     4021 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     4081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6181 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6241 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6301 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6361 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6421 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6481 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6541 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6601 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6661 acgtt
//
