LOCUS       sequence-270259-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G63710 I4_pECIA14 should be used with bait vector 
            AT5G63710 I4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  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 5756)
  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: AT5G63710 I4_pECIA14
FEATURES             Location/Qualifiers
     source          1..5756
                     /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             1052..1075
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2750..2773
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2789..2806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2829..2846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2879..3013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2903..2922)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2957..2976
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3157..3173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3170..3192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3181..3197
                     /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(3205..3211,3212..3229,3230..3235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3250..3271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3388..3405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3559..4147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3639..3658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4318..5109,5110..5178))
                     /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     4941..4960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5179..5283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5351..5369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5407..5429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5529..5548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5742..5756,1..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 tcgtcaacaa ctcaaccaga
      541 tatcgaagga ggagctctgt tgcagctcag agattcgctt aatgattcga gcaatcgtct
      601 aaaatggaca cgcgattttg tgagcccttg ctatagttgg tcttatgtta cctgcagagg
      661 ccagagtgtt gtggctctaa atcttgcctc gagtggattc acaggaacac tctctccagc
      721 tattacaaaa ctgaagttct tggttacctt agagttacag aacaatagtt tatctggtgc
      781 cttaccagat tctcttggga acatggttaa tctacagact ttaaacctat cagtgaatag
      841 tttcagcgga tcgataccag cgagctggag tcagctctcg aatctaaagc acttggatct
      901 ctcatccaat aatttaacag gaagcatccc aacacaattc ttctcaatcc caacattcga
      961 tttttcagga actcagctta tatgcggtaa aagtttgaat cagccttgtt cttcaagttc
     1021 tcgtcttcca gtcacatcct ccaagaaaaa gctggaggtg ctgttccagg gaccaagcgg
     1081 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     1141 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     1201 caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg gctccatcat
     1261 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     1321 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     1381 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     1441 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     1501 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     1561 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     1621 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     1681 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     1741 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     1801 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     1861 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     1921 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     1981 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     2041 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     2101 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     2161 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     2221 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     2281 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     2341 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     2401 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     2461 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     2521 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     2581 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     2641 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     2701 caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg attacaagga
     2761 tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag tttaaacccg
     2821 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2881 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2941 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3001 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3061 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3121 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5281 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5341 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5401 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5461 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5521 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5581 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5641 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5701 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
