LOCUS       sequence-296630-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G21340 N4_pECIA2 should be used with prey vector 
            AT3G21340 N4_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  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 5756)
  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: AT3G21340 N4_pECIA2
FEATURES             Location/Qualifiers
     source          1..5756
                     /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             2009..2032
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2738..2779
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2789..2806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2829..2846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2879..3013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2903..2922)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2957..2976
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3157..3173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3170..3192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3181..3197
                     /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(3205..3211,3212..3229,3230..3235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3250..3271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3388..3405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3559..4147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3639..3658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4318..5109,5110..5178))
                     /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     4941..4960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5179..5283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5351..5369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5407..5429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5529..5548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5742..5756,1..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 agtttggatt gcgggtcatt
      541 gccaaatgag cctccttaca acgatccttc aaccggatta acatactcga cggacgatgg
      601 tttcgtgcag agtggcaaaa ctggaagaat ccagaaagcg ttcgagtcga tcttcagtaa
      661 accgtctttg aagcttagat acttcccgga cggattccga aactgctata ccttgaatgt
      721 cacgcaagac acaaactatc tgatcaaagc tgtatttgtg tatggtaact acgatggtct
      781 taacaatccc ccgagtttcg atctttacct tggtccgaat ctatgggtaa cggttgatat
      841 gaatggacgg accaatggta ctatccagga gattatccac aagaccatat ctaagtctct
      901 ccaggtctgt cttgttaaga caggaacaag ctcacctatg attaatacgt tagagctacg
      961 accacttaaa aacaatactt acaatactca gagtggctct ctgaagtatt tcttccgata
     1021 ttatttcagc ggttcaggcc aaaacatacg gtaccctgat gatgtcaatg atcgtaaatg
     1081 gtatccattc tttgatgcaa aagagtggac agagttaaca accaatctga atataaacag
     1141 ttctaatggt tatgcaccac cagaagttgt gatggcgtca gcctcaacgc ctataagtac
     1201 ttttggaaca tggaacttct catggttatt gccatcttcc acaacccaat tttatgtgta
     1261 catgcatttt gccgagattc aaactctacg gtccctcgat acccgagaat tcaaagtgac
     1321 gttgaatgga aaacttgctt atgaacgcta cagccctaaa acgttagcca ccgaaaccat
     1381 tttctattcg acaccacaac aatgtgaaga tgggacatgc ctcttggagt tgacgaaaac
     1441 acctaagtct actcttcctc ctctcatgaa cgctcttgag gttttcaccg tgatcgattt
     1501 tccacagatg gaaacaaatc cagatgatgt tgctgctatc aagagtatcc aaagcactta
     1561 tggattaagt aaaatcagct ggcaaggaga tccatgcgtt cctaaacagt ttttgtggga
     1621 gggtttaaac tgcaataatc tagataactc cacgccgcct attgtcactt ccttaaactt
     1681 atcgtcaagt catttaacgg ggatcatcgc gcaaggcatt cagaatctga cacacctaca
     1741 agaactagac ttgtcaaata acaatttgac gggaggaata cccgaatttc ttgctgacat
     1801 aaaatcactc ttagtaataa atttaagtgg gaacaatttt aatggctcta ttcctcaaat
     1861 ccttttacag aagaaaggac taaagctaat tcttgaagga aacgccaatc tgatttgtcc
     1921 ggatggatta tgtgtaaaca aagctggcaa tggtggtgcc aagaaaatga atgttgtaat
     1981 accgattgtt gcatcagttg cgtttgtgct ggaggtgctg ttccagggac cagggagcga
     2041 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     2101 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     2161 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     2221 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2281 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     2341 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     2401 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2461 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2521 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2581 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     2641 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     2701 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     2761 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2821 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2881 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2941 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3001 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3061 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3121 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5281 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5341 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5401 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5461 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5521 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5581 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5641 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5701 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
