LOCUS       sequence-275970-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PSKR2 nC10_pECIA14 should be used with bait vector PSKR2
            nC10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  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 7214)
  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: PSKR2 nC10_pECIA14
FEATURES             Location/Qualifiers
     source          1..7214
                     /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             2510..2533
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4208..4231
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4247..4264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4287..4304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4337..4471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4361..4380)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4415..4434
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4615..4631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4615..4631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4628..4650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4639..4655
                     /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(4663..4669,4670..4687,4688..4693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4708..4729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4846..4863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5017..5605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5097..5116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5776..6567,6568..6636))
                     /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     6399..6418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6637..6741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6809..6827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6865..6887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6987..7006
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7200..7214,1..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 agccaaccat gtcatcccaa
      541 cgacttatct gcgctccggg aattggcagg agcgttgaag aacaagtctg ttacagaatc
      601 ttggttaaat ggttcacgtt gttgtgaatg ggatggtgtg ttttgtgaag ggagtgatgt
      661 ttctggtcga gttacaaagt tggttttacc tgaaaaaggt ttggaaggtg tgatttcgaa
      721 gtctttaggg gagttgactg agctgcgagt acttgatcta tctcgtaacc agcttaaagg
      781 cgaagtacca gcggagattt ctaagttaga gcagcttcaa gttcttgatt tgagtcataa
      841 cctgttatca gggtctgttt tgggagtggt ttcgggttta aagctgattc agtcgctgaa
      901 catttcgagc aattcgctta gcgggaagtt atcggatgtt ggagtgtttc ctggtcttgt
      961 gatgcttaat gtaagcaaca atttgtttga gggtgagatt catcctgaac tctgtagctc
     1021 atctggtggg atacaggttc ttgatttatc gatgaatcgt ttggtgggga atcttgatgg
     1081 cttgtacaac tgcagcaaat ctattcaaca gctccatatc gacagcaaca gattgacggg
     1141 ccaacttccg gattatcttt attcgatccg ggagttggag caactatcac tctctggaaa
     1201 ctacttatcc ggagagttaa gcaagaactt gagcaatctc tctggtctga agtctctgtt
     1261 gatatcagag aaccggtttt cggatgtaat tccagatgtt tttggtaacc tcactcaatt
     1321 ggaacacctc gacgtgagct ccaacaagtt ctcgggaagg tttccgccaa gtttatccca
     1381 atgctcgaag ctgcgggttc ttgatcttag gaacaactcg ttatccggtt ctatcaatct
     1441 taacttcact ggatttaccg atctttgcgt gcttgatctc gccagtaatc atttctctgg
     1501 acctcttcct gattcccttg gccactgtcc caagatgaag atcttgagtt tggcgaaaaa
     1561 cgagtttcgc ggcaaaatcc ctgacacctt caagaatctg cagtctctct tgttcctgtc
     1621 cttatccaac aacagctttg tggatttttc tgagacaatg aatgtgctgc aacattgcag
     1681 aaacctctcc actcttattc tctcaaagaa cttcatcggc gaggaaatac caaacaacgt
     1741 cactggtttc gacaacctcg cgattttagc gctaggaaat tgcggtctta gaggtcagat
     1801 tccgagctgg ctattgaact gcaagaagct ggaagttctt gatctctctt ggaatcactt
     1861 ttacggaact atccctcatt ggattggtaa gatggagagt ttgttctaca tagacttctc
     1921 aaacaacact ttgaccggag caatcccggt agccataacc gagctcaaga acctaatccg
     1981 tctaaacgga accgcttctc agatgaccga ctcttctgga attcctctct acgtaaagcg
     2041 gaacaagagc tccaacggtc ttccatataa ccaagtttca agattcccgc catctatcta
     2101 tttgaataat aaccgtctca acgggacgat cttgccagag ataggacgtt tgaaagagct
     2161 tcacatgctg gacttgagca ggaacaactt cactgggacg atacctgatt ccatttcagg
     2221 gcttgacaat ttggaggttc ttgatttatc ttacaatcat ctctacggtt cgattcctct
     2281 gtcttttcag agtctcactt tcttgtcgag gttcagcgta gcgtataacc gtctcactgg
     2341 cgcgattcca tctggaggtc agttctacag cttcccgcac tcaagcttcg aaggaaactt
     2401 aggactttgt cgcgcgattg attctccttg cgatgttctg atgagtaaca tgttgaatcc
     2461 gaaaggttct tcgcgtagga ataacaatgg cggaaagttc gggagaagcc tggaggtgct
     2521 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     2581 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     2641 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     2701 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     2761 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     2821 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     2881 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     2941 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     3001 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     3061 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     3121 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     3181 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     3241 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     3301 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     3361 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     3421 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     3481 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     3541 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     3601 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     3661 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     3721 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     3781 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     3841 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     3901 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     3961 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     4021 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     4081 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     4141 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     4201 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     4261 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     4321 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     4381 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4441 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4501 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4561 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     4621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6781 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6841 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6901 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6961 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7021 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     7081 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7141 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7201 ccagtcacga cgtt
//
