LOCUS       sequence-288215-        6989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G24130 L4_pECIA14 should be used with bait vector 
            AT2G24130 L4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  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 6989)
  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: AT2G24130 L4_pECIA14
FEATURES             Location/Qualifiers
     source          1..6989
                     /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             2285..2308
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3983..4006
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4022..4039
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4062..4079)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4112..4246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4136..4155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4190..4209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4390..4406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4390..4406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4403..4425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4414..4430
                     /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(4438..4444,4445..4462,4463..4468))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4483..4504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4621..4638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4872..4891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5551..6342,6343..6411))
                     /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     6174..6193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6412..6516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6584..6602
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6640..6662)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6762..6781
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6975..6989,1..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 tctaaagaga acgatcatga
      541 gctaataaaa agtgatcaaa tctctttgct ttctttcaaa tcatccattg tttcagatcc
      601 tcaaaactct ctaagcagtt ggatttcttc ttcttcttct tcgtcttcta tgctcgttga
      661 tgtttgtaat tggtctggtg tgaagtgtaa caaagagagc actcaagtaa tcgagcttga
      721 tatcagtggt agagatctcg gcggcgaaat ctcaccgtcg atcgctaatc taaccggttt
      781 aacggttctt gatctttcga gaaacttctt cgttggaaaa atcccacctg agataggttc
      841 attgcatgaa acattgaagc agctaagtct ctctgagaat cttttacatg gaaacattcc
      901 acaagagctt ggtttgctta accgtcttgt ttatctcgat cttggaagca accggttaaa
      961 cggttcaatc ccggttcagc tcttttgcaa cggctcttct tcatctctac aatacataga
     1021 cctttccaac aactctttaa ccggagagat ccctttaaat taccattgtc acctcaaaga
     1081 acttcggttt cttcttctct ggtccaacaa gctcaccggt actgtcccgt cttctctctc
     1141 aaactctacg aatctcaagt ggatggattt ggagtcaaat atgttaagtg gagagttacc
     1201 ttctcaagtc ataagcaaga tgccacaact tcagtttctt tacctttctt ataaccattt
     1261 cgttagccac aacaataaca caaacctcga accttttttc gcctctctcg ccaattcctc
     1321 tgacctacaa gagcttgagt tagccggaaa cagcctcgga ggagagatta cttcttccgt
     1381 aaggcacctc tctgtaaatc ttgtgcagat tcatcttgat caaaaccgta tccatggctc
     1441 tatacctcca gagatatcaa accttttgaa cctaactctt ctaaacttgt cgagtaacct
     1501 cttgtccggt cctatcccgc gtgagctctg taagctaagc aagctcgaaa gggtttatct
     1561 ctcgaataat cacctaaccg gcgaaatccc tatggaactc ggggatattc ctcgccttgg
     1621 tcttcttgat gtttctagaa acaatctttc tggttcaatc cctgatagtt tcggaaacct
     1681 ttctcagctg agaagacttc ttctgtacgg aaaccatctc tctggtactg tccctcaaag
     1741 cctcggcaaa tgtatcaatc ttgagattct tgatctctct cacaacaacc tcacagggac
     1801 catacctgtt gaagttgtct ccaatctcag aaacctgaag ctgtatctca acctctctag
     1861 caaccactta agcggtccaa tacctttgga gctaagcaaa atggatatgg ttctctccgt
     1921 tgatctgtct tccaacgaac tctcagggaa gattccgcct cagctcggaa gctgcattgc
     1981 attagagcat ctgaatcttt caagaaacgg tttctccagt acgcttcctt catcattagg
     2041 ccaacttcca tatctgaaag agctagatgt gtcttttaac agattaaccg gagctatacc
     2101 gccttctttt caacagtctt cgactctcaa gcacttgaat ttctccttca acttgctctc
     2161 tggtaatgta tctgacaaag gatcattctc taagctgact attgagtctt ttcttggaga
     2221 ttctctcttg tgtggctcta taaaaggcat gcaagcctgc aagaagaaac acaagtaccc
     2281 ttctctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     2341 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     2401 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     2461 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     2521 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     2581 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     2641 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     2701 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     2761 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     2821 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     2881 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     2941 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     3001 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     3061 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     3121 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     3181 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     3241 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     3301 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     3361 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     3421 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     3481 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     3541 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     3601 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     3661 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     3721 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     3781 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     3841 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     3901 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     3961 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     4021 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     4081 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     4141 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4201 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4261 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4321 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     4381 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4441 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4501 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4561 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4621 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4681 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5221 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5581 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5641 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5701 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5761 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5821 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5881 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5941 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6001 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6061 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6121 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6181 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6241 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6301 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6361 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6541 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6601 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6661 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6721 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6781 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6841 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6901 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6961 aacgccaggg ttttcccagt cacgacgtt
//
