LOCUS       sequence-296806-        5069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G50230 J8_pECIA2 should be used with prey vector 
            AT3G50230 J8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5069)
  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 5069)
  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: AT3G50230 J8_pECIA2
FEATURES             Location/Qualifiers
     source          1..5069
                     /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             1322..1345
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2051..2092
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2102..2119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2142..2159)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2192..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2216..2235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2270..2289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2470..2486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2470..2486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2483..2505)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2494..2510
                     /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(2518..2524,2525..2542,2543..2548))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2563..2584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2701..2718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2872..3460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2952..2971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3631..4422,4423..4491))
                     /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     4254..4273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4492..4596)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4664..4682
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4720..4742)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4842..4861
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5055..5069,1..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 tctctcccgg cgacgaacta
      541 cttcgactcc ttcttaccat ccgacgccgt cgcacttctc tccttcaaat ccaccgctga
      601 tctcgacaac aagcttctat actctctcac cgagccttac gactactgtc aatggcgcgg
      661 cgtcgattgc tctcaagatc gcgtcgttcg tcttatcctc gacggtgttg gtctccgggg
      721 aagtttctca ccggagactc ttagtcgtct tgatcagctt cgtgttctta gccttgagaa
      781 taactcaatc tcgggttcga ttcctgatct atctcctctc gtcaatctca aaaccctaac
      841 cctaagcaag aatggtttct ccggtactct ttcttcttca attctctctc ttcgtcggtt
      901 aacagaactt gatctatcgt ttaacaactt ctccggtgag attccgtcgg ggatcaatgc
      961 tctgagtcga ctcagttctt taaatctcga gtttaatcga cttaacggaa ctctcccgcc
     1021 gttaaatctt tcttctctga tttcatttaa cgtttccagt aacaacctca ccggtttagt
     1081 tccgttaaca aagactctgt tacgattcaa cgcgtcttcg ttttcgtcaa accctggact
     1141 ctgcggtgag atcattaacc gatcttgcgg tttacattct tcttctccgt tctttggttc
     1201 acctaaacca aacaccacgt catcaacatc ttctgcttct tcgtctgaag caccggttat
     1261 tcaaagtgaa cagaacggtg aagcagcgat gattgttcca ccggtggtta aaaaggttaa
     1321 actggaggtg ctgttccagg gaccagggag cgacaaaact cacacatgcc caccgtgccc
     1381 agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac
     1441 cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga
     1501 ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa
     1561 gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca
     1621 ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc
     1681 ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac
     1741 cctgccccca tcccgggagg agatgaccaa gaaccaggtc agcctgacct gcctggtcaa
     1801 aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa
     1861 ctacaagacc acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct
     1921 caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga
     1981 gggtctgcac aaccactaca cgcagaagag cctctccctg tctccgggta aatctagagg
     2041 gcccttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2101 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     2161 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     2221 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2281 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2341 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2401 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     2461 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4621 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4681 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4741 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4801 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4861 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4921 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4981 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5041 aacgccaggg ttttcccagt cacgacgtt
//
