LOCUS       sequence-227244-        7118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PSKR1 nC5_pECIA14 should be used with bait vector PSKR1 
            nC5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  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 7118)
  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: PSKR1 nC5_pECIA14
FEATURES             Location/Qualifiers
     source          1..7118
                     /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             2414..2437
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4112..4135
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4151..4168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4191..4208)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4241..4375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4265..4284)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4319..4338
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4519..4535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4519..4535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4532..4554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4543..4559
                     /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(4567..4573,4574..4591,4592..4597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4612..4633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4750..4767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4921..5509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5001..5020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5680..6471,6472..6540))
                     /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     6303..6322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6541..6645)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6713..6731
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6769..6791)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6891..6910
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7104..7118,1..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 tcggaatctc agaccacctc
      541 caggtgccat ccacatgacc tcgaagcctt acgtgacttc atagcacatc tcgaaccaaa
      601 accagatggt tggatcaatt cttcttcttc tacagactgc tgcaactgga ccggaatcac
      661 ctgcaattca aacaacaccg gaagagttat tagattggag cttgggaaca aaaagctgtc
      721 ggggaagttg tctgaatctc tcgggaagct agatgagatt agggttctta atctctctcg
      781 aaacttcatc aaagattcga tccctctttc gattttcaac ttgaagaatc tacaaactct
      841 tgatttgagc tctaatgatc tctccggcgg aatcccaaca agtataaatc tcccagctct
      901 gcaaagtttt gatctttctt caaataaatt caatgggtcg cttccgtctc atatctgcca
      961 taactctact caaattaggg ttgtgaaact tgcggtgaac tacttcgccg gaaacttcac
     1021 ttccgggttt gggaaatgtg tcttgcttga gcatctctgt cttggtatga acgatcttac
     1081 tggtaacatc cctgaggatt tgtttcatct caaaagattg aatcttttag ggattcaaga
     1141 gaatcgtctc tctggttcgt tgagtcgtga gattaggaat ctctcaagtc ttgttcgtct
     1201 tgatgtttct tggaatttgt tttccggtga aatccctgat gtgttcgacg aattgcctca
     1261 gttaaagttt ttcttaggtc agaccaatgg attcattgga ggaataccta aatcgttggc
     1321 gaattcaccg agtttgaatc tgcttaactt gaggaacaat tctttatcgg gtcgtttgat
     1381 gttgaattgt acggcgatga ttgctttgaa ctctcttgat ttaggtacca atagattcaa
     1441 tgggaggtta cctgagaatc taccggattg caagcggtta aagaacgtta acctcgcgag
     1501 gaacaccttc catggacaag taccagagag tttcaagaac ttcgagagct tatcttactt
     1561 ctcgttatcg aattcgagtt tggctaatat ctcttcagcg cttgggatac ttcagcattg
     1621 caagaacttg acgactttgg ttcttacatt gaatttccat ggagaggctt tacccgatga
     1681 ttcaagtctt catttcgaga agcttaaggt gcttgtagtg gcgaattgta ggcttactgg
     1741 ttcgatgccg aggtggttaa gctcgagtaa tgaacttcag ttgttggatc tttcttggaa
     1801 ccgtttaacc ggcgctatcc cgagctggat tggtgacttc aaggctctgt tctacttgga
     1861 tttatctaac aactcgttta caggagagat ccctaagagc ttaactaagt tagagagtct
     1921 cactagccgt aatatctcag tcaatgagcc atctcctgat ttcccgttct ttatgaaaag
     1981 aaacgagagc gcgagagcgt tgcaatacaa tcagattttc gggttcccgc caacaattga
     2041 gcttggtcat aacaatctct ctggacctat ttgggaggag tttggtaatc tgaagaagct
     2101 tcatgtgttt gatttgaaat ggaatgcatt atctggatca atacctagct cgctttctgg
     2161 tatgacgagc ttggaagctc tcgatctctc taataaccgt ctttcgggtt cgatcccggt
     2221 ttctctgcaa cagctctcgt ttctgtcgaa gttcagtgtt gcttataaca atctctcggg
     2281 agtaatacct tccggtggtc agtttcagac gtttccaaac tcgagctttg agagtaacca
     2341 tctctgcggg gaacacagat tcccctgttc tgaaggtact gagagtgcat tgatcaaacg
     2401 gtcaagaaga agcctggagg tgctgttcca gggaccaagc ggcaatcccg gcggcgatct
     2461 ggccccccag atgctgcgcg agctgcagga gaccaatgcc gccctgcagg acgtgcgcga
     2521 gctgctgcgc caccgcgtga aggagatcac cttcctgaag aacaccgtga tggagtgcga
     2581 cgcctgcggc atgcagccct ctagaggctc cggctccatc atcccagttg aggaggagaa
     2641 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc
     2701 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac
     2761 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc
     2821 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca
     2881 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca
     2941 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga
     3001 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac
     3061 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg
     3121 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac
     3181 gcagctcatc tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt
     3241 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct
     3301 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg
     3361 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct
     3421 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct
     3481 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct
     3541 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact
     3601 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga
     3661 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc
     3721 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta
     3781 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga
     3841 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga
     3901 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct
     3961 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct
     4021 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc
     4081 gggttctaga ggaagcacct ccggcgataa cgattacaag gatgatgatg ataaggccag
     4141 cggcaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     4201 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     4261 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4321 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4381 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     4441 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     4501 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4621 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4681 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4741 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4801 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5341 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6661 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6721 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6781 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6841 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6901 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6961 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     7021 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     7081 aagttgggta acgccagggt tttcccagtc acgacgtt
//
