LOCUS       sequence-237788-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G66830 X005_pECIA2 should be used with prey vector 
            AT1G66830 X005_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  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 5114)
  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: AT1G66830 X005_pECIA2
FEATURES             Location/Qualifiers
     source          1..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1812..1835
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2541..2582
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2592..2609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2632..2649)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2682..2816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2706..2725)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2760..2779
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2960..2976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2960..2976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2973..2995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2984..3000
                     /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(3008..3014,3015..3032,3033..3038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3053..3074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3191..3208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3362..3950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3442..3461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4121..4912,4913..4981))
                     /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     4744..4763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4982..5086)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtctct aaacgatcaa ggtcttgctc ttctttcctt taaacaatcc atacaaaacc
     1021 agagtgactc tgtttttacc aattggaact cttcagattc aaacccttgt tcttggcaag
     1081 gtgttacatg taattatgat atgagagttg tctcgattcg tcttcctaat aaacgtcttt
     1141 cgggttctct tgatccatcc attggaagtc tcttgtccct tcgtcacatt aacctaagag
     1201 acaatgactt tcaagggaag ttacctgtgg aactctttgg acttaaaggg cttcaaagct
     1261 tggtgctttc tggaaattct ttttccgggt ttgtcccaga agagattggt agcttgaaga
     1321 gtctaatgac tctggatctt tcggagaatt ctttcaacgg ttcgatttct ttgtcgctta
     1381 ttccatgcaa gaagctgaaa acgcttgttc ttagcaagaa cagtttctct ggagatcttc
     1441 caacagggtt aggttctaac ttggttcatc ttcggacact gaatctttct ttcaatcgct
     1501 tgactggaac gatccccgaa gatgtaggaa gtctggagaa tctgaaaggt acacttgatc
     1561 tttctcataa cttcttctca ggtatgattc caacaagtct tggaaatctc cctgagcttc
     1621 tctatgtcga tctttcttac aataatctca gtggtccgat accaaaattc aacgttctgt
     1681 tgaatgccgg tccaaatgct ttccaaggga accctttcct ctgtggatta ccgataaaga
     1741 tttcttgcag cacaagaaac acacaagtag ttccttctca actatataca agaagagcca
     1801 accatcattc tctggaggtg ctgttccagg gaccagggag cgacaaaact cacacatgcc
     1861 caccgtgccc agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac
     1921 ccaaggacac cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga
     1981 gccacgaaga ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg
     2041 ccaagacaaa gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca
     2101 ccgtcctgca ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag
     2161 ccctcccagc ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac
     2221 aggtgtacac cctgccccca tcccgggagg agatgaccaa gaaccaggtc agcctgacct
     2281 gcctggtcaa aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc
     2341 cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc ttcttcctct
     2401 acagcaagct caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg
     2461 tgatgcatga gggtctgcac aaccactaca cgcagaagag cctctccctg tctccgggta
     2521 aatctagagg gcccttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     2581 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     2641 tgccttctaa ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat
     2701 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2761 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2821 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2881 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg
     2941 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3781 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5101 aagtgccacc tgac
//
