LOCUS       sequence-297025-        6050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G29730 X066_pECIA2 should be used with prey vector 
            AT1G29730 X066_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6050)
  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 6050)
  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: AT1G29730 X066_pECIA2
FEATURES             Location/Qualifiers
     source          1..6050
                     /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             2303..2326
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3032..3073
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3083..3100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3123..3140)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3173..3307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3197..3216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3251..3270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3451..3467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3451..3467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3464..3486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3475..3491
                     /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(3499..3505,3506..3523,3524..3529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3544..3565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3682..3699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3853..4441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3933..3952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4612..5403,5404..5472))
                     /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     5235..5254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5473..5577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5645..5663
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5701..5723)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5823..5842
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6036..6050,1..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 tctccttctc tgcacccaga
      541 tgaagtggaa gcgctgaagg atatcactga aacattgggt gtgaagcact tgaacctaag
      601 tgaagatcca tgtctcacaa agactctagt gatatctcaa ggtgttctca aggaaggaca
      661 gaacagcaca atcagatgtg actgtcattt caataactac agcacttgtc atatcaaaca
      721 ctttgtcctc cagaaattca atcttccagg tagacttcct ccaatgctgt acaagtttcg
      781 gcatcttgaa tcgattgact tatacaataa ttacctttat ggctcaattc ctatggaatg
      841 ggcctcactg ccttacctca aatctatctc tgtctgtgca aaccgcttat caggagacat
      901 tcccaaagga ttggggaagt ttattaacct caccctttta gttcttgaag ccaatcagtt
      961 ttcgggaact attcctaagg aactagggaa cctagtgaat ctacaaggac taggactctc
     1021 ttccaaccaa cttgtcggag gcctccccaa gacattagca aaactaacaa agctaactaa
     1081 tctccattta agtgataatc gcctgaacgg atcgattcca gagtttattg ggaagttacc
     1141 aaaacttcaa agattagaac tctatgcgag tggccttaga ggacctattc cagactccat
     1201 ttttcatctg gagaatttga tcgacgtgag aatcagtgac acggttgcag gattgggaca
     1261 tgttcctcag ataactagca cgagcctgaa atatttggtt ttgagaaaca taaacttatc
     1321 tggaccaatt ccaaccagta tctgggatct gcccagttta atgactctgg atctttcctt
     1381 taataggctg actggagaaa taccagcata tgcaactgcc ccaaaataca catatttagc
     1441 tggaaacatg ttgtctggaa aggttgaaac aggagctttc cttactgcaa gcacaaatat
     1501 tgatctttcc tataataatt tcacgtggtc tccaatgtgc aaagagagga agaatataaa
     1561 cacatatgag agctcacact caaaaaaccg tctaaccaga cttcttccat gttccgccat
     1621 aaagcaatgc caaaattata gtagatcgct gcatataaac tgtgggggac ctgatgtaac
     1681 catcgaaaac tctcgaggaa ggtttctgta tgaaggtgat aactatggac ttaccggatc
     1741 agctacgaac tattatagga aaaattgggg atacagcaat actggtgact ttatggatga
     1801 tgcaataaca gaagatacat acacagtttc atcagaatct gcagtttcag caaaatatcc
     1861 agacctttac cagaatgctc ggcgctctcc cctcagtttg gcttactatg cattttgctt
     1921 cgaaaatgga agctacaatg tgaaactcca ttttgcagag attcagttct cagatgtgga
     1981 accatacact aaactagcga aacgggtttt taacatttac attcagggga agttgatttg
     2041 ggaggatttc agcattagag aggaggctaa tggaactcac aaagaagtta taagagaagt
     2101 aaacacgacc gtgactgata acactttaga gatacggctt tactgggcag ggaaaggcac
     2161 aatgatcatt cctcaaagag ggtactatgg ctctcttatc tctgcagtct cagtttgtcc
     2221 cagttcagaa tctgaatgcg gtggtatgaa gaagaaaatc tcaaagctta agggacctga
     2281 tctaagaaca ggttccttca gtctggaggt gctgttccag ggaccaggga gcgacaaaac
     2341 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     2401 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     2461 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     2521 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     2581 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     2641 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     2701 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     2761 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     2821 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     2881 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     2941 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     3001 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     3061 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     3121 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3181 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3241 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3301 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3361 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3421 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5581 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5641 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5701 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5761 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5821 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5881 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5941 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6001 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
