LOCUS       sequence-296648-        5846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G48740 O3_pECIA2 should be used with prey vector 
            AT5G48740 O3_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5846)
  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 5846)
  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: AT5G48740 O3_pECIA2
FEATURES             Location/Qualifiers
     source          1..5846
                     /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             2099..2122
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2828..2869
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2879..2896
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2919..2936)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2969..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2993..3012)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3047..3066
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3247..3263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3247..3263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3260..3282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3271..3287
                     /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(3295..3301,3302..3319,3320..3325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3340..3361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3478..3495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3649..4237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3729..3748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4408..5199,5200..5268))
                     /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     5031..5050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5269..5373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5441..5459
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5497..5519)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5619..5638
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5832..5846,1..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 agtccagatg gtttcttgag
      541 cttatcatgt ggcggctcaa gttacaccgc tgcttataac atctcatggg tttcagacaa
      601 cgattatatc gaaacaggta acactaccac ggttacttac gccgagggga attctacttc
      661 cagtgttccc atcagattat tcccggatcc tcaaggtcgc caatgttaca agttacctgt
      721 gaggaaagac ttgtcttcag ttctgattcg tgctacattc gtttatagaa actatgatag
      781 ccagaatagt cctccagcat ttcatgtctc tcttggaagg agaatcacta gtactgttga
      841 tcttagaacc aatgatcctt ggattgaaga gttagtttgg ccagtcaaca atgattctct
      901 tttgctttgc ttgcttgcgg ttaaaggaag aggtattccg gttatctctt ctctagaagt
      961 acggccactt cctttaggtt cttacaaata cagcctggag ggttcaccag atataattct
     1021 caggagaagt taccgaatca atagcggata cacaaatggg actatccggt atccttcaga
     1081 tccatttgat cgaatatggg atccggatca aagttattca ccgtttcacg cctcttggag
     1141 cttcaatgga ctcacaaaat tgaattcctt caatattact gagaatccac cagcttctgt
     1201 tcttaagaca gcaagaattc ttgctcgcaa agaaagtctt tcatatactc tttctctaca
     1261 tacaccaggg gactactaca ttatcctcta ctttgctggg atactctcac tctccccttc
     1321 ttttagtgtg acgatcaatg atgaagtcaa acaatctgat tacaccgtaa ctagctcaga
     1381 agctggtacc ttgtacttta cgcaaaaagg aatcagtaag ttaaacatca ccctcagaaa
     1441 aatcaagttc aatcctcaag tgagtgctct tgaggtatat gaaatcctcc aaatccctcc
     1501 tgaagcttct tctaccacag tttcggcact taaagtcata gagcaattta ccgggcaaga
     1561 ccttggatgg caagacgacc catgcacacc gcttccttgg aaccacatcg aatgtgaagg
     1621 aaaccgtgtt acatcattgt tcctttcaaa aatcaacctg aggtccatta gtccaacatt
     1681 cggagacttg ctcgatctca aaacactcga cttgcataat acatcactta caggagcaat
     1741 acagaatgta ggcagcctta aggatctcca gaaactgaat ttgagcttca atcagttaga
     1801 atcttttggc tccgagctgg aagatctggt taaccttgaa gttctggacc tgcaaaacaa
     1861 cagcttacaa ggatcagttc ctgaaacatt aggaaagctg aagaaacttc gtctcctgaa
     1921 tctggaaaac aataacttag taggtccctt gccacagtct cttaacataa cgggtctgga
     1981 agtcaggata acgggaaatc catgcctttc gttctcttca atatcatgca acaatgtgtc
     2041 ttcaacaatt gacacaccac aagtcactat ccctatcaat aagaaacaaa gaaaacagct
     2101 ggaggtgctg ttccagggac cagggagcga caaaactcac acatgcccac cgtgcccagc
     2161 acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct
     2221 catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc
     2281 tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc
     2341 gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca
     2401 ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc
     2461 catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg tgtacaccct
     2521 gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg
     2581 cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta
     2641 caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac
     2701 cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggg
     2761 tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaat ctagagggcc
     2821 cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     2881 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     2941 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3001 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3061 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3181 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     3241 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3301 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3361 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3421 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3481 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3541 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3601 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3661 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3721 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3781 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3841 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3901 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3961 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4021 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4081 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4141 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4201 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4261 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4321 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4381 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5341 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5401 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5461 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5521 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5581 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5641 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5701 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5761 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5821 gccagggttt tcccagtcac gacgtt
//
