LOCUS       sequence-296822-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G25320 X002_pECIA2 should be used with prey vector 
            AT1G25320 X002_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5150)
  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 5150)
  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: AT1G25320 X002_pECIA2
FEATURES             Location/Qualifiers
     source          1..5150
                     /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             1403..1426
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2132..2173
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2183..2200
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2223..2240)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2273..2407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2297..2316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2351..2370
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2551..2567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2564..2586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2575..2591
                     /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(2599..2605,2606..2623,2624..2629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2644..2665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2782..2799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2953..3541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3033..3052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3712..4503,4504..4572))
                     /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     4335..4354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4573..4677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4745..4763
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4801..4823)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4923..4942
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5136..5150,1..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 aatgctctaa acgacgaagg
      541 gtttgctctt ttaactctga agcagtctat ttcaaaagat ccagatggtt ctttaagcaa
      601 ttggaactca gagaatcaaa acccttgttc ttggaatgga gtcacttgtg atgataacaa
      661 ggttgttgtc tctcttagca tcccaaagaa gaaactttta ggttatcttc cttcttctct
      721 tggtttactc tctaatctcc gtcatttgaa tttgagaagc aatgagctta gtgggaactt
      781 acctgttgag ctttttaagg ctcaaggtct tcaaagtttg gttctttatg ggaatttctt
      841 gtctgggtcc attccaaatg agattggtga cttaaagttt ctgcaaattt tggatctttc
      901 tcgtaattct cttaatgggt ctattccaga atcggttttg aagtgtaata ggcttaggag
      961 ctttgatttg agtcagaata atctcactgg ttctgttcct agtggatttg gtcagtcttt
     1021 ggcttctttg cagaagcttg atctttcttc taacaatctt attggtcttg ttcctgatga
     1081 tttggggaat ttgactagat tgcaaggaac tcttgatttg tctcataatt cgtttagtgg
     1141 ctcgattcca gctagtttgg ggaatttgcc tgagaaagtt tatgtcaatc tagcttacaa
     1201 caacttgagt ggaccaatcc cacaaaccgg tgctttggtg aacagaggac caactgcatt
     1261 cttggggaat ccaagactct gtggccctcc attgaaggat ccttgtttgc cagatacgga
     1321 tagttcttca acttctcacc cttttgtacc ggataacaac gagcaaggtg gaggagggtc
     1381 aaagaaagga gaaggtttaa gtctggaggt gctgttccag ggaccaggga gcgacaaaac
     1441 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     1501 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     1561 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     1621 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     1681 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     1741 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     1801 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     1861 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     1921 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     1981 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     2041 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     2101 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     2161 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     2221 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     2281 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2341 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2401 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2461 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     2521 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2701 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4681 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4741 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4801 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4861 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4921 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4981 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5041 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5101 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
