LOCUS       sequence-298863-        6032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G24130 L4_pECIA2 should be used with prey vector 
            AT2G24130 L4_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6032
                     /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             3014..3055
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3065..3082
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3105..3122)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3155..3289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3179..3198)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3233..3252
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3433..3449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3433..3449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3446..3468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3457..3473
                     /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(3481..3487,3488..3505,3506..3511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3526..3547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3664..3681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3835..4423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3915..3934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4594..5385,5386..5454))
                     /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     5217..5236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5455..5559)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5627..5645
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5683..5705)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5805..5824
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6018..6032,1..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 agggaccagg gagcgacaaa actcacacat gcccaccgtg
     2341 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     2401 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     2461 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     2521 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     2581 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     2641 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     2701 caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt
     2761 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     2821 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     2881 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     2941 tgagggtctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatctag
     3001 agggcccttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     3061 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3121 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3181 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3241 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3301 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3361 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     3421 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3481 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3541 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3601 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3661 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3721 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3781 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3841 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3901 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3961 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4021 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4081 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4141 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4201 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4261 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4321 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4381 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4441 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4501 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4561 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4621 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4681 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4741 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4801 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4861 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4921 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4981 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5041 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5101 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5161 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5221 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5281 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5341 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5401 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5461 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5521 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5581 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5641 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5761 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5821 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5881 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5941 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6001 ggtaacgcca gggttttccc agtcacgacg tt
//
