LOCUS       sequence-296627-        5696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G28990 N1_pECIA2 should be used with prey vector 
            AT2G28990 N1_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5696)
  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 5696)
  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: AT2G28990 N1_pECIA2
FEATURES             Location/Qualifiers
     source          1..5696
                     /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             1949..1972
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2678..2719
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2729..2746
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2769..2786)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2819..2953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2843..2862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2897..2916
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3097..3113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3097..3113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3110..3132)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3121..3137
                     /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(3145..3151,3152..3169,3170..3175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3190..3211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3328..3345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3499..4087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3579..3598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4258..5049,5050..5118))
                     /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     4881..4900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5119..5223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5291..5309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5347..5369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5469..5488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5682..5696,1..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 agtttagatt gtggattacc
      541 tagtgacgaa tctccttacg atgattcgtt taacggatta acattcacat cggattccac
      601 tttcatccaa acaggaaaaa ttgatagtgt cgataaagat cttaacataa acttatcaaa
      661 acaatatttg actcttagat acttcccaga aggaaaacgt aattgttata gtcttgatgt
      721 caagcgtggc acaacttatc tcatcgtggt tagctttgta tatggaaatt atgatggtct
      781 taaccgtgat ccaaacttcg atatccatct tggtcccaat aagtggaaaa gaatagattt
      841 ggatggagag aaagaaggta caagagaaga gatcatacac aaagccaggt caaactcttt
      901 ggatatttgt cttgttaaga caggagaaac tttgccaata atttcagcca tagaaatacg
      961 gccactaagg aataatacgt atgttacaca atctggttcg ctgatgatgt ctttccgggt
     1021 ttatcttagc aattcagatg catctataag gtatgctgat gacgtccatg atcgtatttg
     1081 gtctccattc aacggatctt cacatacaca tataacaact gatctcaata taaacaactc
     1141 aaatgcatat gagattccaa aaaatatact ccaaactgct gccataccaa gaaacgccag
     1201 tgcgccgctg attattacgt gggatccact acccattaat gcggaggtat acttgtacat
     1261 gcactttgcg gagattcaaa cccttgaagc caacgagacg aggcagtttg acgttatctt
     1321 gagaggaaac tttaatcatt caggatttag tccaaccaag ttaaaggtat ttacattata
     1381 cacagaagaa ccaatgaaat gtggttcaga aggttgttat ttacagctag taaaaacacc
     1441 gaactcaact ctgccacctc tgatcaacgc tattgaggca tacagtgtta tcgagttctc
     1501 acaattggaa acaagtctta gtgatgtgga cgctatcaag aacatcaaaa atacttataa
     1561 gctgaataaa atcacttggc aaggagatcc atgtctccct caagatttgt cgtgggaaag
     1621 tattagatgc acctacgtag atggctccac ttcaccaaca atcatttcct tagatttgtc
     1681 gaaaagtgga ttaaatggga gtatacccca aattttacaa aatttcacgc aattacaaga
     1741 actggactta tcaaataata gcttgactgg accggtgcct atcttccttg ccaacatgaa
     1801 aacgttatct ttaataaact taagtgggaa caatctcagt ggttcagttc ctcaagctct
     1861 tcttgacaaa gaaaaagaag gacttgtgtt aaaacttgaa ggaaatccag atttgtgcaa
     1921 atccagtttt tgcaataccg aaaagaaact ggaggtgctg ttccagggac cagggagcga
     1981 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     2041 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     2101 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     2161 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2221 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     2281 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     2341 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2401 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2461 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2521 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     2581 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     2641 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     2701 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2761 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2821 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2881 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2941 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3001 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3061 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3121 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3181 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3241 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3301 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3361 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3421 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3901 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4141 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4201 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4561 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4861 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5221 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5281 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5341 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5401 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5461 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5521 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5581 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5641 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
