LOCUS       sequence-297096-        5330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector MRLK J10_pECIA2 should be used with prey vector MRLK 
            J10_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5330)
  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 5330)
  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: MRLK J10_pECIA2
FEATURES             Location/Qualifiers
     source          1..5330
                     /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             1583..1606
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2312..2353
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2363..2380
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2403..2420)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2453..2587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2477..2496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2531..2550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2731..2747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2731..2747)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2744..2766)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2755..2771
                     /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(2779..2785,2786..2803,2804..2809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2824..2845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2962..2979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3133..3721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3213..3232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3892..4683,4684..4752))
                     /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     4515..4534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4753..4857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4925..4943
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4981..5003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5103..5122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5316..5330,1..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 ggtgtagtga taacacaagc
      541 tgattaccaa ggtcttcaag cagttaaaca agaactgatt gatccaagag gcttcttgag
      601 aagctggaac ggttctggat tcagtgcttg ctctggaggc tgggctggaa taaaatgtgc
      661 tcaaggtcaa gtcatcgtca ttcaacttcc atggaagagt cttggaggta gaatctctga
      721 gaaaatcgga cagcttcaag ctcttcgtaa gcttagtctt catgacaaca accttggagg
      781 ttcgattcca atgtcattgg gactcattcc taatcttaga ggagttcagc ttttcaacaa
      841 ccgtctcact ggctcaatcc ctgcttctct cggtgtatca catttccttc aaacgcttga
      901 tctcagcaat aacttgctct ctgagattat tccaccgaat cttgctgact cttctaagct
      961 tcttaggctt aatctcagct ttaattcact gtctggtcaa atcccagtga gtctctctcg
     1021 gtcttcttcc cttcagtttc ttgctcttga ccataacaac ctctctggtc caatcttaga
     1081 tacttggggt agtaagtctc ttaatctccg tgtcttatct cttgatcaca actctctctc
     1141 tggtccgttc ccattttcac tctgcaactt aactcagctg caagattttt cctttagtca
     1201 taataggatc cgtggaaccc taccctctga gctaagcaaa ctcactaagc ttagaaaaat
     1261 ggatattagt ggtaacagtg ttagtggcca tatccctgaa accctaggga acatttcttc
     1321 tcttatacat ttggatttgt ctcaaaacaa actcaccgga gagattccta tttcgatttc
     1381 tgacctggaa agcctgaact tcttcaatgt ctcttataac aacttatctg gtcctgttcc
     1441 tactctcttg tcccaaaagt tcaattcctc ttcttttgtc gggaactcac tgctttgcgg
     1501 atacagcgtt tcgacaccat gtcctactct tccttcaccg tctcctgaaa aagaaaggaa
     1561 accatcccac aggaatctga gtctggaggt gctgttccag ggaccaggga gcgacaaaac
     1621 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     1681 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     1741 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     1801 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     1861 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     1921 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     1981 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     2041 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     2101 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     2161 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     2221 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     2281 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     2341 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     2401 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     2461 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2521 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2581 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2641 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     2701 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2761 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2821 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2881 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2941 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3001 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3061 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3121 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3181 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3241 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3301 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3361 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3421 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3481 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3541 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3601 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3661 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3721 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3781 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3841 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3901 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3961 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4021 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4081 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4141 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4201 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4261 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4321 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4381 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4441 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4501 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4561 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4621 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4681 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4741 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4801 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4861 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4921 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5101 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5161 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5221 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5281 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
