LOCUS       sequence-274073-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G36180 X019_pECIA14 should be used with bait vector 
            AT4G36180 X019_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  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 7400)
  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: AT4G36180 X019_pECIA14
FEATURES             Location/Qualifiers
     source          1..7400
                     /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             2696..2719
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4394..4417
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4433..4450
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4473..4490)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4523..4657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4547..4566)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4601..4620
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4801..4817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4801..4817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4814..4836)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4825..4841
                     /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(4849..4855,4856..4873,4874..4879))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4894..4915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5032..5049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5203..5791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5283..5302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5962..6753,6754..6822))
                     /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     6585..6604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6823..6927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6995..7013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7051..7073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7173..7192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7386..7400,1..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 tctcaagccg agattgatgc
      541 tctcacggcg tttaaactca acctccatga ccctctcggt gctttaactt catgggatcc
      601 ttcaacacca gcagctccat gtgactggcg tggcgttggc tgtactaacc accgtgtcac
      661 tgagatccgt cttcctcgtc tccaactctc cggacgaatc tccgatcgta tctccggtct
      721 acgcatgcta cggaagctca gtctccggtc taactctttc aacggtacta tccctacttc
      781 actcgcttac tgcacgcgcc tactctccgt ctttcttcag tacaactcac tctccggtaa
      841 actacctccg gcgatgagga atctcacctc gcttgaggtc tttaacgtcg ctggaaaccg
      901 tctctccggt gagatccccg ttggtttacc atcgagtctc cagttccttg acatttcgtc
      961 gaacaccttc tctggtcaga taccgagtgg actcgctaac ttgactcagc ttcagcttct
     1021 taatctctcg tataatcagt taaccggcga gattccggcg agtctaggta acctccaaag
     1081 tcttcagtac ttatggcttg atttcaatct cttgcaagga acattaccgt ctgctatctc
     1141 aaactgctcc tccttagttc acttgagcgc gtcggagaac gaaatcggcg gcgtgattcc
     1201 cgccgcgtac ggtgctcttc cgaaactgga ggttctctcg ctctccaata acaatttttc
     1261 cggtactgtg ccgttttctc tcttctgcaa cacatcgctt acgattgttc agttagggtt
     1321 caatgctttc tccgatatcg tacggcctga gacgacggcg aactgtcgta ccggtttaca
     1381 ggtcttggat ctccaggaaa atcgaatctc cggtcgtttt ccgttgtggt tgactaatat
     1441 tttgtcacta aagaatcttg atgtttctgg aaacttgttt tccggtgaga ttccgccgga
     1501 tattggtaac ttgaagagat tagaggaact gaaattggct aataactctc ttactggtga
     1561 gattcctgtt gagattaagc aatgtggttc gttagatgtt ttagatttcg aaggaaatag
     1621 tttaaaaggt cagattccag aatttcttgg ctacatgaag gctttaaagg ttttatcttt
     1681 aggcagaaac agcttctcag gttatgttcc ttcgtctatg gtgaatctgc agcagcttga
     1741 gaggctaaac ttaggtgaaa acaacttaaa tgggagtttt ccggtggagt tgatggcgtt
     1801 gacaagtctg tctgagctgg atctgagtgg aaatcggttt tccggggcag ttccggttag
     1861 tatcagcaac ttaagtaacc ttagtttttt gaatttgagt gggaatggat tttctggtga
     1921 gataccagct agtgtaggca atctgtttaa gttaacagct cttgatttga gcaagcagaa
     1981 tatgtctggt gaggttccgg ttgagctctc tggtttgccc aacgtgcaag tgattgcatt
     2041 gcagggaaac aacttctctg gggttgtacc tgaagggttt agtagcttgg tgagtttaag
     2101 gtatgtgaac cttagctcga actcattctc cggggaaata ccgcagactt ttgggtttct
     2161 tcggttgttg gtttctttgt cactgtcaga taatcacatc tctgggtcta ttccacctga
     2221 aattggaaac tgctcagctc ttgaagttct tgagttaaga tctaaccggc taatgggtca
     2281 tattccagct gatctctctc gtcttccgcg tttgaaggtg cttgacttgg gtcagaacaa
     2341 cttgtcaggc gaaatcccac cagaaatctc tcagagctcg tctctcaact cattgtcact
     2401 tgaccacaat catctttcag gagtcatacc tggatctttt tctggtttat caaacttgac
     2461 aaagatggat ctttctgtta ataacttaac cggggagatt ccggctagcc tcgcccttat
     2521 ttctagtaac ctggtatact tcaacgtctc aagcaacaac cttaaaggag agattccagc
     2581 ttctttaggc tcaagaatca acaacacatc agaattttca ggcaacacag agctctgcgg
     2641 gaagccattg aacagaagat gtgaaagcag cacagcggag gggaagaaga agaaactgga
     2701 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     2761 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     2821 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     2881 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     2941 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     3001 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     3061 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     3121 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     3181 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     3241 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     3301 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     3361 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     3421 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     3481 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     3541 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     3601 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     3661 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     3721 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     3781 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     3841 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     3901 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     3961 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     4021 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     4081 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     4141 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     4201 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     4261 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     4321 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     4381 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     4441 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     4501 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     4561 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4621 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4681 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     4741 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     4801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5641 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6961 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     7021 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     7081 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     7141 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     7201 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     7261 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     7321 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     7381 gttttcccag tcacgacgtt
//
