LOCUS       sequence-296998-        5228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G14390 O9_pECIA2 should be used with prey vector 
            AT1G14390 O9_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5228)
  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 5228)
  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: AT1G14390 O9_pECIA2
FEATURES             Location/Qualifiers
     source          1..5228
                     /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             1481..1504
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2210..2251
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2261..2278
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2301..2318)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2351..2485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2375..2394)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2429..2448
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2629..2645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2629..2645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2642..2664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2653..2669
                     /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(2677..2683,2684..2701,2702..2707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2722..2743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2860..2877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /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     4413..4432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4651..4755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4823..4841
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4879..4901)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5001..5020
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5214..5228,1..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 agtgaatcaa gaactctatt
      541 agaaatccaa aagcatttac aatatccacc aactcttcga tcatggagca actggaccaa
      601 tttctgctac ctcccttctt ctccttcttt caaaatcctc tgtttcaatg gtcatgtaac
      661 tgaattaacc gtcacgggga acagaaccgt taagctcccc ggaagattct caagcgactc
      721 actattcact gttcttacaa aactctcaaa cttaaagaca ttgtctctgg tctctcttgg
      781 catctctggt cctctccctt cacagatcat caggttatct tcatctcttc aatctctgaa
      841 tctcagttct aatttcatct ctgggaatat tcctaaagag atctcgtcgt tgaagaatct
      901 gagaagtctt gttctagcga acaatttgtt taacggaagt gttcctgatc tcagaggatt
      961 gtcgaatctt caagagctga atttaggcgg taataaactc ggtcccgaag ttgttccttc
     1021 acttgcaagt aacttgatta ctatttcatt gaagaacaac tcttttggat caaagattcc
     1081 agaacagatc aagaagctga ataagcttca gagcttagat ctttcctcca acaagttcac
     1141 tggctcaatc ccaagattcc tgctttcgct tccttcgctt cagaatctaa gtttagcaca
     1201 aaacttgctg agtggatcac ttccaaattc atcgttgtgc aactccaagc ttagaatttt
     1261 agatgtttct cggaatcttc tgactgggaa gcttccatct tgcttctctt ccaagaaaca
     1321 gacagtactg ctcttcacat tcaattgctt gtctataaat ggttctcctt ctgcaaagta
     1381 tcagcgtcca gttaccttct gtgaaaacga agcaaagcaa gcagtagctg ctgtgaagtc
     1441 tgacactaaa gataaagaaa ggaaagaaga agatacagga ctggaggtgc tgttccaggg
     1501 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     1561 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     1621 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     1681 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     1741 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     1801 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     1861 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     1921 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     1981 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     2041 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     2101 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     2161 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     2221 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     2281 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     2341 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2401 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2461 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2521 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     2581 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     2641 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2701 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2821 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4801 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4861 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4921 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4981 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5041 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5101 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5161 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5221 acgacgtt
//
