LOCUS       sequence-297049-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G49780 X065_pECIA2 should be used with prey vector 
            AT5G49780 X065_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  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 5516)
  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: AT5G49780 X065_pECIA2
FEATURES             Location/Qualifiers
     source          1..5516
                     /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             1769..1792
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2498..2539
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2549..2566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2589..2606)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2639..2773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2663..2682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2717..2736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2917..2933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2917..2933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2930..2952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2941..2957
                     /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(2965..2971,2972..2989,2990..2995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3010..3031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3148..3165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3319..3907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3399..3418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4078..4869,4870..4938))
                     /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     4701..4720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4939..5043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5111..5129
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5167..5189)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5289..5308
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5502..5516,1..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 tccttgaatt caaataaatt
      541 taatggaacg attccggctt ccattggcct gttatcgaaa ctatattggt ttgatatagc
      601 tgacaatcag atagaaggaa agcttccagt ttctgatggg gcttctttac ctggacttga
      661 tatgcttctt caaactaagc attttcattt tggaaaaaac aagctttctg gtgatatccc
      721 tgaaaagcta ttcagtgcca acatgacttt aaaacatttg ctattcgatg gaaacctact
      781 aaccggtgaa atcccacaaa gcctcagcct tgttaaaaca ttgacggtgt tacgccttga
      841 taggaataga ctaagtggag aaattcctcc aagtcttaat aatctcacaa atcttcaaga
      901 actgtacttg tccgacaaca aatttaccgg tagtcttcca agtttaacca gcttgaccag
      961 tctctccaca ttagctgtca gcaataaccg attaacttcc tcacaaattt catcatggat
     1021 ctcattatta ccgacttcct tggcaacatt aaggatggca gggctccaac ttcaaggtcc
     1081 aataccaacc tccctcttca gtcttcctga attgcagact gttatcctaa agcgtaactg
     1141 gctaaacgaa acactagact tcggtaccaa caagagccaa aacctagatt ttgtggattt
     1201 acaatataat gacataacag agtatattaa acaaccagct aataaaggaa gcagcaggat
     1261 agtaatcttg gcaaataatc cagtgtgccc cgaggtaggg aacccaccaa atgaatactg
     1321 catagaagtc gaacacaatt cctcatattc ttcccctaag aacacatgtg gtcgttgtag
     1381 tggtgaggac agggaaccca ttcccaccac gtgccgttgt gtgtatccaa tcacaggaac
     1441 actcactttt aggtcacctt ccttctcagg gtattccaac aacgatacat tcgagaacct
     1501 caggctaaac ctaactggtt tctttgagaa tagaaattat acagtggatt ctgtggccat
     1561 aagaaacata agagaagatg aagatgatca ttatcttcta atagatctct ctctctttcc
     1621 atataaacaa gacagattta atgagacggg aatggatagt gttatttcca ggtttagcac
     1681 acagacttat aagcctccta atacctttgg cccttacata ttcaaagcca ataagtacaa
     1741 taagttccct gcaggaggtt caaactcgct ggaggtgctg ttccagggac cagggagcga
     1801 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     1861 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     1921 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     1981 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2041 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     2101 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     2161 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2221 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2281 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2341 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     2401 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     2461 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     2521 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2581 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2641 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2701 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2761 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2821 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     2881 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2941 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3001 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3061 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3121 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3721 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5041 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5101 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5161 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5221 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5281 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5341 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5401 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5461 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
