LOCUS       sequence-270119-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G51890 M5_pECIA14 should be used with bait vector 
            AT1G51890 M5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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 6644)
  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: AT1G51890 M5_pECIA14
FEATURES             Location/Qualifiers
     source          1..6644
                     /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             1940..1963
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3638..3661
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3677..3694
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3717..3734)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3767..3901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3791..3810)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3845..3864
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4045..4061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4045..4061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4058..4080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4069..4085
                     /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(4093..4099,4100..4117,4118..4123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4138..4159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4276..4293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4447..5035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4527..4546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5206..5997,5998..6066))
                     /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     5829..5848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6067..6171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6239..6257
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6295..6317)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6417..6436
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6630..6644,1..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 tcaggattca taagcttaga
      541 ttgtggtttg gtgcctacgg aaattactta tgtggaaaag tcgacgaata taacatacag
      601 atcagacgca acttacatcg acagtggagt tcccgggaag atcaatgaag tgtacagaac
      661 acagtttcag caacaaattt gggccttgag aagcttccct gagggtcaaa gaaattgtta
      721 caacttcagt ctcaccgcaa aacgtaagta tctaatcaga ggaaccttta tctatgggaa
      781 ttatgacggt ttgaatcaac tcccgagctt tgatctttac atcggtccaa acaaatggac
      841 ctctgtttcg atccccggag tgagaaatgg ttcagtctcc gagatgatcc atgtcttaag
      901 acaagaccat cttcaaattt gtcttgtgaa aacaggagaa actacaccgt ttatttcttc
      961 attggaactt cgtcctttga acaataatac atacgtcaca aaaagtggat cgcttattgt
     1021 ggtcgcaaga ctttactttt cacccactcc accatttctc aggtatgatg aggacgtcca
     1081 tgaccgaatt tggattccat tcttagataa caaaaattcc ttgttaagca cggaactctc
     1141 tgttgataca agtaacttct acaatgtgcc tcaaactgta gcgaaaactg ctgctgtccc
     1201 tttaaatgct actcagcctc tgaaaataaa ttggagtctc gacgacatca cttcacagtc
     1261 atatatatac atgcatttcg ctgaaatcga gaatcttgaa gctaatgaga ccagagaatt
     1321 caatattact tacaatggtg gcgaaaattg gttctcctat tttagacctc caaagtttcg
     1381 tataacaact gtatacaatc cagcagctgt gagttctcta gatgggaatt tcaacttcac
     1441 tttctcgatg accggtaact ctactcatcc tcctcttatc aacggccttg agatttatca
     1501 agttctagag cttccacagc ttgatacata ccaagatgaa gtttccgcta tgatgaatat
     1561 caagacaata tatggattga gcaaaaggtc tagctggcaa ggagatccat gtgctcctga
     1621 gttatataga tgggaaggtt taaactgtag ttatccaaac tttgcgccac cgcagatcat
     1681 atccttgaac ttgagtggaa gcaatttgag tggtaccata acatctgata tatccaagct
     1741 aacacatttg agagaactag atttatcaaa caatgactta tcaggagata ttccatttgt
     1801 tttttctgat atgaagaatt tgacactcat aaacttgagt ggaaacaaga atctaaatcg
     1861 ctcagttcca gagactcttc agaagaggat agataacaaa tctttaacac taattagaga
     1921 tgaaaccgga aaaaatagtc tggaggtgct gttccaggga ccaagcggca atcccggcgg
     1981 cgatctggcc ccccagatgc tgcgcgagct gcaggagacc aatgccgccc tgcaggacgt
     2041 gcgcgagctg ctgcgccacc gcgtgaagga gatcaccttc ctgaagaaca ccgtgatgga
     2101 gtgcgacgcc tgcggcatgc agccctctag aggctccggc tccatcatcc cagttgagga
     2161 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     2221 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     2281 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     2341 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     2401 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     2461 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     2521 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     2581 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     2641 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     2701 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtggag gccgaaagta
     2761 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     2821 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     2881 tgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     2941 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     3001 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     3061 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     3121 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     3181 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     3241 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     3301 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     3361 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     3421 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     3481 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     3541 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     3601 gcacccgggt tctagaggaa gcacctccgg cgataacgat tacaaggatg atgatgataa
     3661 ggccagcggc accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3721 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3781 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3841 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3901 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3961 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     4021 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4081 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4141 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4201 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4261 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4321 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4381 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4441 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4501 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4561 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4621 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4681 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4741 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4801 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4861 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4921 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4981 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5041 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5101 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5161 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5221 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5281 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5341 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5401 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5461 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5521 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5581 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5641 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5701 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5761 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5821 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5881 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5941 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6001 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6061 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6121 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6181 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6241 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6301 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6361 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6421 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6481 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6541 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6601 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
