LOCUS       sequence-296842-        5990 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector AT5G10020 X022_pECIA2 should be used with prey vector 
            AT5G10020 X022_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  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 5990)
  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: AT5G10020 X022_pECIA2
FEATURES             Location/Qualifiers
     source          1..5990
                     /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             2243..2266
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2972..3013
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3023..3040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3063..3080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3113..3247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3137..3156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3191..3210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3391..3407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3391..3407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3404..3426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3415..3431
                     /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(3439..3445,3446..3463,3464..3469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3484..3505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3622..3639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3793..4381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3873..3892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4552..5343,5344..5412))
                     /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     5175..5194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5413..5517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5585..5603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5641..5663)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5763..5782
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5976..5990,1..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 accgagacgg agctccgatc
      541 tctactcgag ttccgtaaag ggatccgtga cgagacctca caccaacgaa tctcatggtc
      601 agatacaagc agcctcactg atccatctac ttgccctaat gactggcctg gtatctcttg
      661 cgaccctgaa accggttcaa tcatcgccat taatctcgac cgtcgtggtc tctccggcga
      721 actcaaattc agcactctct ctggtcttac tcgtcttcgt aacctctctt tatccggaaa
      781 cagcttctcg ggtcgggtcg ttccttcatt gggtggaatc tcgtcgcttc aacacttaga
      841 tctgtcggat aacggtttct acggacctat accgggtcgg atctcggagc tttggagttt
      901 gaatcatctc aatttgtctt ccaacaagtt cgaaggtggg tttcctagtg gctttcggaa
      961 tcttcagcag ctaagatctt tggatttgca taagaacgag atttggggag atgttggtga
     1021 gattttcact gagcttaaga atgttgagtt cgttgatttg agctgtaacc gtttcaacgg
     1081 tggcttatct ttaccaatgg agaacatttc cagcatttcc aacactctcc gtcacttgaa
     1141 tttgagccac aatgcgttaa atggcaagtt tttcagtgag gagtccattg gttcgttcaa
     1201 gaatctcgag attgttgact tagagaataa ccagatcaat ggtgagttac cgcattttgg
     1261 atcacagccg agtttgagaa tattgaagct tgctcgtaac gagttgttcg gtttagtacc
     1321 tcaagagttg ttgcagagtt cgattccttt gctagagcta gatctaagtc gaaacggttt
     1381 tacaggatca attagtgaaa tcaattcatc aacgttaact atgttgaatc tttcttctaa
     1441 cggcctgtct ggggatcttc cgtcgagttt taaaagttgc tcagtgatag atttgagtgg
     1501 gaatactttc tctggcgacg tgtctgttgt tcaaaagtgg gaagctactc cagatgttct
     1561 tgatcttagc tcaaacaact tgtcgggaag cttgccgaac tttacttctg cattttcaag
     1621 attaagtgtg ttgagcatta ggaataattc tgtctctggt agcttgcctt ctctgtgggg
     1681 tgattctcaa ttctctgtga ttgatctcag ttcaaataag ttcagtggct tcattccagt
     1741 gagtttcttc acttttgcaa gcttgagaag tttgaatctt tctaggaata acttggaggg
     1801 gccaattcct tttcgaggtt cacgtgcaag cgaacttttg gttctaaatt cttatccgca
     1861 gatggagtta cttgaccttt caaccaattc tttgacgggt atgctgcctg gggatatagg
     1921 tacaatggaa aaaattaaag tgctgaatct tgccaataat aagctgtccg gggaacttcc
     1981 aagcgacctc aacaagctta gtggcctttt gttccttgac ttgtcaaaca atactttcaa
     2041 gggtcaaatt cccaataagc ttccttccca aatggtggga ttcaatgtgt cgtataacga
     2101 cctatcaggc ataattccgg aagaccttag aagctacccg ccttcttcat tttatcctgg
     2161 taactccaag ctaagtctcc caggacggat tccagcggat tcttctggtg atctgtcttt
     2221 acctgggaaa aaacatcatt cgctggaggt gctgttccag ggaccaggga gcgacaaaac
     2281 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     2341 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     2401 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     2461 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     2521 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     2581 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     2641 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     2701 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     2761 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     2821 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     2881 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     2941 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     3001 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     3061 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3121 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3181 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3241 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3301 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3361 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3541 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3601 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4201 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5521 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5581 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5641 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5701 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5761 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5821 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5881 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5941 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
