LOCUS       sequence-238708-        4853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BIR4 C2_pECIA2 should be used with prey vector BIR4 
            C2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4853)
  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 4853)
  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: BIR4 C2_pECIA2
FEATURES             Location/Qualifiers
     source          1..4853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1551..1574
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2280..2321
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2331..2348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2371..2388)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2421..2555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2445..2464)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2499..2518
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2699..2715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2699..2715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2712..2734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2723..2739
                     /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(2747..2753,2754..2771,2772..2777))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2792..2813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2930..2947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3101..3689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3181..3200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3860..4651,4652..4720))
                     /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     4483..4502
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4721..4825)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtctca cgcagaagac gatgtactct gtctcaaagg ctttaaatca tcactcaaag
     1021 atccttcaaa tcaactcaac acatggtctt tccctaattc atcatcgtcg atttgcaaac
     1081 tcaccggagt ttcttgctgg aacgctaaag agaatcgaat tctctctctt cagcttcaat
     1141 caatgcaact ctctggtcaa atccctgaat ctcttaaact atgtcggagt ttacaatctt
     1201 tagatctttc ctttaatgat ttctccggtt tgattccttc acaaatatgt tcttggcttc
     1261 cttatcttgt tactttagat ctttccggta acaaactctc cggttcgata ccgtctcaga
     1321 tcgttgactg taaattctta aacagcttag ctctaaacca gaacaagtta accggttcga
     1381 ttccgtctga attaacccgg ttaaaccgtc tccaacgact ttctctagct gataacgatc
     1441 tctctggttc gattccctcc gagctatctc attacggaga agatggtttt cgcggtaatg
     1501 gcggactttg tggtaagccg ttatccaact gcggttcgtt taacggtaaa ctggaggtgc
     1561 tgttccaggg accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac
     1621 tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct
     1681 cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca
     1741 agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg
     1801 agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc
     1861 tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga
     1921 aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat
     1981 cccgggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc
     2041 ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca
     2101 cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca
     2161 agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca
     2221 accactacac gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag
     2281 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     2341 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc
     2401 gctgatcagc ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa
     2461 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2521 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2581 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     2641 gtggtatggc tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt
     2701 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2761 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2821 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3541 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4501 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4561 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4621 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4681 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4741 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4801 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
