LOCUS       sequence-270426-        6134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RLK X023_pECIA14 should be used with bait vector RLK 
            X023_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6134)
  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 6134)
  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: RLK X023_pECIA14
FEATURES             Location/Qualifiers
     source          1..6134
                     /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             1430..1453
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3128..3151
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3167..3184
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3207..3224)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3257..3391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3281..3300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3335..3354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3535..3551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3535..3551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3548..3570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3559..3575
                     /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(3583..3589,3590..3607,3608..3613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3628..3649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3766..3783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3937..4525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4017..4036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4696..5487,5488..5556))
                     /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     5319..5338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5557..5661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5729..5747
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5785..5807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5907..5926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6120..6134,1..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 accacagacg gtgttcttct
      541 tctctctttc cgttactcaa tcgttgacga tcctctttac gtttttcgga gctggagatt
      601 cgacgacgag actccttgct cttggcgtgg tgtcacgtgc gatgcatctt cccggcacgt
      661 gactgttctg tcacttccaa gctcgaacct taccggcaca ctaccttcaa atttgggttc
      721 actcaattca cttcaaagac ttgatctttc caacaattcc atcaatgggt ctttcccggt
      781 ttcgcttctc aacgcgacgg agcttcgatt tcttgatctg tccgataatc acatctccgg
      841 tgcactaccg gcgagttttg gcgcgctttc gaacctccaa gtgttgaatc tctccgataa
      901 ttccttcgtc ggcgaattac cgaacacatt aggatggaac cggaacttaa cggagatttc
      961 acttcagaaa aactatttat ccggcgggat tccgggaggt tttaagtcga cggagtatct
     1021 tgatctctcg tcaaatttga tcaaaggctc gttgccgtca catttcagag ggaatcgtct
     1081 acgctatttc aacgcttcgt acaacagaat ctccggcgag attccgtcag gtttcgccga
     1141 cgaaatcccg gaagacgcca ccgttgatct ctcattcaac caacttacag gtcaaatccc
     1201 gggttttcgg gttctcgata accaagaatc caactctttc tccggtaacc cgggtctctg
     1261 cggatccgac catgcaaaac acccttgtcg tgacggtgaa gcaacctctc cacctccatc
     1321 gccgactcca aattctcctc ctgcattagc tgctatacca aatactattg gcttaaccaa
     1381 tcacccaatt agctccaaaa ccggtccgaa atcaaaatgg gatcataaac tggaggtgct
     1441 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     1501 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     1561 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     1621 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     1681 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     1741 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     1801 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     1861 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     1921 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     1981 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     2041 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     2101 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     2161 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     2221 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     2281 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     2341 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     2401 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     2461 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     2521 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     2581 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     2641 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     2701 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     2761 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     2821 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     2881 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     2941 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     3001 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     3061 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     3121 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     3181 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     3241 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3361 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3421 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3481 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     3541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3601 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5701 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5761 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5821 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5881 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5941 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6001 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6061 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6121 ccagtcacga cgtt
//
