LOCUS       sequence-298807-        6530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G35710 X075_pECIA2 should be used with prey vector 
            AT1G35710 X075_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  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 6530)
  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: AT1G35710 X075_pECIA2
FEATURES             Location/Qualifiers
     source          1..6530
                     /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             2783..2806
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3512..3553
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3563..3580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3603..3620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3653..3787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3677..3696)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3731..3750
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3931..3947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3931..3947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3944..3966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3955..3971
                     /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(3979..3985,3986..4003,4004..4009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4024..4045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4162..4179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4333..4921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4413..4432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5092..5883,5884..5952))
                     /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     5715..5734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5953..6057)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6125..6143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6181..6203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6303..6322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6516..6530,1..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 tctgcaacta tcgcagaagc
      541 aaatgctctt ctgaaatgga aatccacttt cacgaactca tcaaaactgt cttcatgggt
      601 ccatgatgca aacaccaata ctagcttttc ctgcaccagt tggtatggtg tttcttgcaa
      661 ctcacgaggg agtatcgaag agttaaacct cactaacact ggtatagaag gtactttcca
      721 agatttcccc ttcatttctc tctcaaacct tgcttacgta gatcttagca tgaacctttt
      781 atctggaacc atccctcctc aattcggaaa cctctctaaa ctcatctact ttgacctctc
      841 cacaaatcac ttaaccggag aaatctctcc ttccttagga aatctcaaga acttgacggt
      901 actttatctt caccaaaact atctaactag tgtcattccc tcggaactag gcaacatgga
      961 atccatgaca gatttagcgc taagtcagaa caaacttacc ggttcaattc cttcttcctt
     1021 aggaaatctc aagaacttga tggttcttta tctttacgag aactatctaa ctggtgtcat
     1081 tcccccggaa ctaggtaaca tggaatccat gacagattta gctttaagtc agaacaaact
     1141 taccggttca attccttcta ccttaggaaa tctcaagaac ttgatggttc tttatcttta
     1201 cgagaactat ctaactggtg tcattccccc agaaataggc aacatggaat ccatgacaaa
     1261 tttagcgcta agtcagaaca aacttactgg ttcaattcct tcttccttag gtaatctcaa
     1321 gaacttgacg cttctttctc ttttccagaa ctatctaact ggtggcattc ccccgaaact
     1381 aggcaacata gaatccatga tcgatttaga gttgagtaac aacaaactta ccggttcaat
     1441 tccttcttcc ttaggaaatc tcaagaattt gactattctt tatctttacg agaactattt
     1501 aactggtgtc attcccccgg aactaggcaa catggaatcg atgattgatt tacagttgaa
     1561 taacaacaaa cttaccggtt caattccttc ttccttcgga aatctcaaga acttgacgta
     1621 tctttatctt tacctgaatt atttaactgg tgtcattccc caggagctag gcaacatgga
     1681 atccatgatc aatttagatt taagtcagaa caaacttaca ggttctgttc ctgattcctt
     1741 tggtaacttt accaagctgg aaagtttgta ccttcgtgtt aaccacctct ctggtgcgat
     1801 cccgccagga gtagcaaact cttcacatct aactaccttg atacttgaca ccaacaactt
     1861 cactggtttc tttcctgaaa ctgtttgcaa aggtaggaag cttcaaaata tctcattgga
     1921 ttacaatcac ctcgaaggtc ctatcccgaa aagcttgaga gattgcaaga gcctgatcag
     1981 agcaagattt ctaggaaaca aattcactgg cgatattttt gaagcttttg ggatttaccc
     2041 agaccttaac ttcattgatt tcagccacaa caaattccac ggtgagattt ctagcaactg
     2101 ggagaagagt ccaaagctag gtgccttgat catgtcaaac aacaacatca cgggtgctat
     2161 cccaacagag atttggaaca tgacacaact agttgagctg gatctatcta ccaacaacct
     2221 ctttggtgaa cttccagaag caattggaaa tctcacgaat ttgtcgaggc tacgacttaa
     2281 tgggaatcag ttatctggaa gagttcctgc aggtttaagt ttcttaacca atcttgagtc
     2341 tcttgactta tcctcaaaca atttcagctc agagatccca caaacctttg actccttttt
     2401 aaagcttcat gacatgaacc tgagcagaaa caaattcgac ggaagcattc cccgactatc
     2461 aaagctgact cagttaacgc aacttgatct cagccacaac cagctcgacg gagaaatccc
     2521 atcacaacta agctccctgc aaagcctcga caagctcgac ctctcacaca acaatctctc
     2581 aggtctcatt ccaacaactt tcgagggcat gatagcgctg acgaacgtcg atatatcaaa
     2641 caataaactc gaaggtccgc ttccagatac tccaacgttt cgaaaagcaa cagcagatgc
     2701 attggaggaa aacataggct tatgcagtaa tattcctaaa caaagattga aaccatgccg
     2761 tgagttgaag aaaccgaaga aactggaggt gctgttccag ggaccaggga gcgacaaaac
     2821 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     2881 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     2941 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     3001 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     3061 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     3121 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     3181 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     3241 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     3301 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     3361 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     3421 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     3481 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     3541 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     3601 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3661 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3721 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3781 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3841 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3901 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5161 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5281 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5521 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5581 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5761 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5941 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6061 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     6181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     6301 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6361 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     6421 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6481 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
