LOCUS       sequence-296821-        5870 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector AT2G27060 B10_pECIA2 should be used with prey vector 
            AT2G27060 B10_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  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 5870)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AT2G27060 B10_pECIA2
FEATURES             Location/Qualifiers
     source          1..5870
                     /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             2123..2146
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2852..2893
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2903..2920
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2943..2960)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2993..3127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3071..3090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3271..3287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3271..3287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3284..3306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3295..3311
                     /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(3319..3325,3326..3343,3344..3349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3364..3385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3502..3519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3673..4261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3753..3772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4432..5223,5224..5292))
                     /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     5055..5074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5293..5397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5465..5483
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5521..5543)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5643..5662
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5856..5870,1..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 tctgactttg aagctctttt
      541 ggagcttaag aaaggttttc agggtgaccc ttctcgaaaa gttcttactt catgggatgc
      601 taaagcttta tcctctgata ggtgtccgtt aaactggtac ggagttactt gtagtagcgg
      661 aggtgttact tctattgatt tgaatggttt cggtctgctt gggagtttca gttttccagt
      721 cattgttggt ctgagaatgc ttcagaattt gtcgattgcg aataatcagt ttagtggaac
      781 tctctctaac attggttcgt taacgtctct taagtacttg gatgtttcgg gtaacttgtt
      841 ccatggcgca ttaccttctg gaattgagaa tttgaggaac cttgagtttg tgaatctctc
      901 tggtaacaac aacttgggag gagtgattcc ttctggtttt gggagtcttg ccaagttgaa
      961 gtaccttgat ttgcaaggca acagtttctc tggggaagtg atgagtctgt tttcacagct
     1021 aattagtgtg gagtatgtag atataagcag gaacaacttc tcgggttcgc ttgatctggg
     1081 tctagcaaag tcaagctttg tttcctcgat tcgtcatttg aacgtaagtg ggaattcttt
     1141 ggtaggggag ctatttgctc atgatggtat cccctttttc gatagcttag aggtgtttga
     1201 tgctagcagc aatcaattgt ctggttctgt tccagtcttc agctttgttg tctctctcaa
     1261 gattcttcgg ttacaagaca atcagttgtc ggcttcttta ccaccgggcc ttctgcaaga
     1321 gagttcaacc atcttaactg accttgatct tagcctcaat caacttgaag gtcctatcgg
     1381 aagtataacc tcttcaacac tagagaagct caatttgtct tcaaatagac tatcgggatc
     1441 attacctttg aaagttggac attgtgccat tattgatctg agtaacaata agatatcagg
     1501 agaattgtca aggatacaga attggggaga ttctgtagaa attattcggt tgagttcaaa
     1561 ctcactcact ggaacactac ctggccaaac ttctcaattc ctgaggctga cttcactcaa
     1621 ggctgctaac aactcgttgc agggtgtact accatttatt ttaggaacgt atcctgagct
     1681 gaaagagatt gatctcagtc acaaccagct tagtggcgtc ataccgagta atctgttcat
     1741 atctgcaaag ttgacggaac tcaacttgtc aaacaataat ttctctggtt ctcttcctct
     1801 tcaagatgca agtactgtgg gaaaccttag cttgaccaat attggtctgt ctcataactc
     1861 gttaggtgga gttctctcag aggaactaac tcgcttccac aatctgattt ctcttgacct
     1921 ttcttataac aactttgaag ggaatatccc tgatggcctt ccagacagcc taaagatgtt
     1981 cactgtctct gccaataatc tctctggaaa tgtgccggag aatcttagac ggtttccaga
     2041 ttcagcgttt catccaggga acgctttact gaatgtcccc atttctctac caaaagataa
     2101 aacagatata accttacgga agctggaggt gctgttccag ggaccaggga gcgacaaaac
     2161 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     2221 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     2281 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     2341 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     2401 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     2461 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     2521 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     2581 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     2641 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     2701 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     2761 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     2821 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     2881 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     2941 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3181 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3241 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3301 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3361 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3421 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3841 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4081 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5401 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5461 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5521 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5581 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5641 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5701 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5761 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5821 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
