LOCUS       sequence-296644-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT4G29450 O1_pECIA2 should be used with prey vector 
            AT4G29450 O1_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  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 5729)
  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: AT4G29450 O1_pECIA2
FEATURES             Location/Qualifiers
     source          1..5729
                     /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"
     RBS             550..558
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1982..2005
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2711..2752
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2802..2819)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2852..2986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2876..2895)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2930..2949
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3130..3146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3130..3146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3143..3165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3154..3170
                     /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(3178..3184,3185..3202,3203..3208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3223..3244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3361..3378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3532..4120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3612..3631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4291..5082,5083..5151))
                     /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     4914..4933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5152..5256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5324..5342
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5380..5402)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5502..5521
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5715..5729,1..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 tcaggttaca tcagcatcga
      541 ttgtggcata cctccttacg atacccctga agacactatg actaacatta attacgtctc
      601 cgatgaagct ttcataacca ccggcgtaaa cttcaaggtc tctgaagaat acggctatcc
      661 aaaaaacccg gtattgctgt caactcttgc tgaagtaaga gctttccctc aaggaaacag
      721 aaactgttac accttaaaac tttcacaagg aaaagatcat ctttatctca ttagagcttc
      781 ctttatgtat ggaaactacg atggtaaaaa agctttgcct gagtttgatc tctatgtaaa
      841 cgtgaacttc tggtcgacag tgaaattcaa aaatgcttct gatcaagtca ccaaagagat
      901 tttaagcttt gcagagtcgg atacaatata tgtttgcctt gtgaataaag gcaaaggaac
      961 tccttttatt tcagggttag agctccgacc agtgaacagc tcaatctacg gtactgaatt
     1021 tggaagaaat gtctctttgg ttctctatcg aagatgggac attggatatt taaacggaac
     1081 aggacggtac caagacgata gattcgatcg tatttggtct ccatactcct caaatatctc
     1141 ctggaactcg attataacat cgggctatat cgatgttttt cagaacggtt attgtccacc
     1201 tgatgaagtt attaagacag ctgctgctcc tgaaaacgtt gatgatccgt tggaattgtt
     1261 ctggacatca gacgatccaa atgttcggtt ctacgcatat ctttacttcg cggaactcga
     1321 aacactagag aagaatgaga ctagaaagat caagattttg tggaatggtt cccctgtttc
     1381 tgaaacttct ttcgaacctt cttcaaaata ttcaacgacg ttttctaatc caagagcttt
     1441 cactggtaaa gatcactgga tttctattca gaagactgta gattcaacgc ttccaccgat
     1501 tctcaatgct atagagattt ttaccgcaca atctctagat gaattttcca ccacaattga
     1561 agacattcat gctatagaaa gcataaaagc aacttataaa gtgaacaagg tttggagtgg
     1621 tgatccttgc tccccaagat tattcccttg ggaaggtgtt ggatgcagcg acaacaataa
     1681 taatcatcaa atcaagtccc tgaatcttag ctcaagcgga ttactaggac caatagtctt
     1741 ggcatttaga aatctctctc ttcttgagtc attggattta tccaacaatg acctgcaaca
     1801 aaatgtgcct gagtttttgg ctgatctaaa acatttgaaa gtcctgaatt tgaaaggaaa
     1861 caattttact gggtttatcc caaaatccct tatgaaaaag ttaaaggccg gtttacttac
     1921 gctcagtgcg gatgagcaaa acctttgtaa ctcatgtcaa gaaaagaaga agaagaagag
     1981 cctggaggtg ctgttccagg gaccagggag cgacaaaact cacacatgcc caccgtgccc
     2041 agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac
     2101 cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga
     2161 ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa
     2221 gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca
     2281 ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc
     2341 ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac
     2401 cctgccccca tcccgggagg agatgaccaa gaaccaggtc agcctgacct gcctggtcaa
     2461 aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa
     2521 ctacaagacc acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct
     2581 caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga
     2641 gggtctgcac aaccactaca cgcagaagag cctctccctg tctccgggta aatctagagg
     2701 gcccttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2761 tcatcgtcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     2821 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     2881 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2941 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3001 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3061 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     3121 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3181 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5281 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5341 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5401 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5461 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5521 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5581 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5641 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5701 aacgccaggg ttttcccagt cacgacgtt
//
