LOCUS       sequence-238704-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector PRK5 I7_pECIA2 should be used with prey vector PRK5 
            I7_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  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 4991)
  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: PRK5 I7_pECIA2
FEATURES             Location/Qualifiers
     source          1..4991
                     /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             1689..1712
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2418..2459
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2469..2486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2509..2526)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2559..2693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2583..2602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2637..2656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2837..2853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2837..2853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2850..2872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2861..2877
                     /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(2885..2891,2892..2909,2910..2915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2930..2951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3068..3085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3239..3827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3319..3338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3998..4789,4790..4858))
                     /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     4621..4640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4859..4963)
                     /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 ggtggttgtg ccggattcag atgcagattg tctcttgcga ttcaaagata
     1021 cattagcaaa tggttcagaa tttcgtagtt gggatccatt atcatcacca tgtcaaggaa
     1081 atacagcaaa ttggtttggt gttctttgta gcaattacgt ttggggatta caacttgagg
     1141 gaatgggttt gaccgggaaa ctaaacctcg acccattggt tccaatgaag aatctgcgaa
     1201 ccataagctt catgaacaat aacttcaatg gcccaatgcc tcaagttaag aggtttactt
     1261 cgctgaaatc tttgtatttg tctaataacc ggttttcggg ggagataccc gcagatgcat
     1321 ttctaggtat gccacttttg aagaaaattt tgctggctaa taacgcattc cgaggaacaa
     1381 ttccttcttc tctagcctct ttgccaatgc ttttagagtt gaggctaaat ggtaatcaat
     1441 ttcaagggca aataccatct tttcaacaga aggatcttaa attagctagc tttgaaaaca
     1501 atgatctcga tggaccgata cctgaaagtc ttcgaaacat ggatccaggc tcttttgcag
     1561 gtaacaaagg tttgtgtgat gctcccttaa gtccatgttc ctcttcctct ccgggggttc
     1621 ctgttgttcc ggtatctccc gttgatccaa aatctacttc tcctcccacg gggaaaaagg
     1681 ccggatctct ggaggtgctg ttccagggac cagggagcga caaaactcac acatgcccac
     1741 cgtgcccagc acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca
     1801 aggacaccct catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc
     1861 acgaagaccc tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca
     1921 agacaaagcc gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg
     1981 tcctgcacca ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc
     2041 tcccagcccc catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg
     2101 tgtacaccct gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc
     2161 tggtcaaagg cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg
     2221 agaacaacta caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca
     2281 gcaagctcac cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga
     2341 tgcatgaggg tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaat
     2401 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     2461 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     2521 cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca
     2581 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2641 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2701 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2761 agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa
     2821 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3001 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3061 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3961 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4021 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4081 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4141 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4201 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4261 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4321 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4381 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4441 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4501 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4561 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4621 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4681 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4741 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4801 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4861 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4921 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4981 tgccacctga c
//
