LOCUS       sequence-297004-        5336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G03770 O12_pECIA2 should be used with prey vector 
            AT3G03770 O12_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5336)
  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 5336)
  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: AT3G03770 O12_pECIA2
FEATURES             Location/Qualifiers
     source          1..5336
                     /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             1589..1612
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2318..2359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2369..2386
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2409..2426)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2459..2593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2483..2502)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2537..2556
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2737..2753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2737..2753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2750..2772)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2761..2777
                     /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(2785..2791,2792..2809,2810..2815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2830..2851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2968..2985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3139..3727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3219..3238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3898..4689,4690..4758))
                     /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     4521..4540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4759..4863)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4931..4949
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4987..5009)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5109..5128
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5322..5336,1..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 tctcaaaccc tcttgaggct
      541 gcaacagctt ctctactatc ccaaagtgtt aaacagctgg aacaactata cagatttctg
      601 caattctgag ccaagccctt cgctaaccgt tgtgtgttat gaagatagtg taacgcagct
      661 tcacattata ggagacaatg gaactcatat gttaccaaag agtttctcaa taaactcgtt
      721 tgttactact cttgttaagc tccctgatgt taaagttctc acgtttgtgt cccttgggtt
      781 atggggttgg ttacctcaga aaatcaaccg tttatcctct ttggagattc tcaatgtgag
      841 ttccaatttc ctctttggac caatccctca tgaactctcg tcgcttgcta ctcttcagac
      901 gcttatactc gacgagaata tgttttctgg tgagttaccg gattggattg attctttgcc
      961 gagtttagct gtgttgagct tgagaaagaa cgtgcttaat gggtcattgc cttcttcttt
     1021 gagtagcttg tcaggtctta gagttcttgc tttagctaat aaccggttta atggagcttt
     1081 gcctgattta agccatttga caaaccttca agtgcttgat cttgaaggaa actcttttgg
     1141 accgttgttt cctcggttaa gtaacaagct ggttactctt atactcagca agaacaagtt
     1201 tagatctgca gtttcagctg aagaagtaag ttctctgtat cagcttcaac atttggatct
     1261 ttcctacaat acattcgttg gtccatttcc tacttccttg atgtctcttc ccgcgataac
     1321 ttacttgaac atttcacaca acaaactcac cgggcggctt tcagcaaatc tttcttgcaa
     1381 ctcacagctg atgtttgtag atatgtcatc aaatcttctt accggaagtt tgcctacttg
     1441 ccttaaaccg agttccggca ctagcagaga tgttgtctat gcaagcaact gtttggctac
     1501 tacgaatgaa gatcaacgtc cagtttcttt ttgtagtaat gaagctcttg ctgttggaat
     1561 cctaccacag aggaggaaca aagtttccct ggaggtgctg ttccagggac cagggagcga
     1621 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     1681 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     1741 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     1801 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     1861 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     1921 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     1981 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2041 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2101 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2161 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     2221 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     2281 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     2341 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2401 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2461 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2521 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2581 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2641 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     2701 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2761 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2821 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2881 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4201 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4501 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4861 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4921 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4981 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5041 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5101 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5161 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5221 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5281 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
