LOCUS       sequence-269961-        6689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector FRK1/SIRK M10_pECIA14 should be used with bait vector 
            FRK1/SIRK M10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  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 6689)
  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: FRK1/SIRK M10_pECIA14
FEATURES             Location/Qualifiers
     source          1..6689
                     /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             1985..2008
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3683..3706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3722..3739
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3762..3779)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3812..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3890..3909
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4090..4106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4090..4106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4103..4125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4114..4130
                     /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(4138..4144,4145..4162,4163..4168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4183..4204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4321..4338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4492..5080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4572..4591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5251..6042,6043..6111))
                     /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     5874..5893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6112..6216)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6284..6302
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6340..6362)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6462..6481
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6675..6689,1..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 tctggtttca taagtataga
      541 ttgcgggata ccggatgatt ctagctacaa cgatgagact acaggtataa agtatgtttc
      601 ggattcggcg tttgttgatt caggaacaac aaagagaatt gcagctcagt ttcaatcaag
      661 tggttttgat agacacttgt tgaacgtgag aagtttccct caaagcaaga gaagctgtta
      721 cgatgttccg acgccgagag gcaaaggttt taagtatcta atcagaactc gtttcatgta
      781 cgggaactat gatgatcttg gaagagtacc cgagttcgat ctctatctag gagtaaactt
      841 ttgggactct gttaaactcg acgatgcaac aactatactc aacaaagaga taatcaccat
      901 tccactttta gacaatgttc aagtgtgtgt tgttgataag aacgcaggaa ctccattttt
      961 gtctgtcttg gagatacggc tcttgttgaa cactacttat gagactcctt acgatgcact
     1021 taccctcctt cggagattag attacagcaa aacgggaaaa cttccatcga ggtacaaaga
     1081 tgacatatat gaccgtatat ggacaccgcg tatagtgagt tcagaataca agatattaaa
     1141 tacatctctc actgtcgatc aatttcttaa caatggctac caaccggctt ctactgtcat
     1201 gagcactgca gaaacagcgc gaaacgagag cctctaccta acgcttagtt tcagaccgcc
     1261 cgaccctaac gcaaagtttt atgtatacat gcactttgct gaaattgaag tactgaaaag
     1321 caaccagacg agagaattca gcatctggct aaatgaggat gtaatctctc cttcgtttaa
     1381 gcttcggtac ttgcttaccg acacattcgt tacaccggat cccgtgagcg gaattaccat
     1441 taacttctct cttcttcaac cacctggtga atttgtactt ccaccgatca ttaacgccct
     1501 tgaggtctat caagtcaatg agttccttca gatcccaact catccacagg atgttgatgc
     1561 catgaggaag attaaagcca cgtatagagt gaagaagaac tggcaaggag atccttgtgt
     1621 tcccgtagat tattcttggg aaggtattga ctgtatccaa agtgataaca ctactaatcc
     1681 tagagtcgtt tcactaaata tatcttttag tgaattacga ggccagatag atccagcctt
     1741 ctccaacctt acatctataa gaaaattaga tttatccggt aatactttaa caggagaaat
     1801 acctgctttc ctcgctaact taccaaactt gaccgaatta aacgttgaag gaaacaagtt
     1861 aacgggcata gttccacaaa gattgcatga aagatcaaag aatggatctc tttccttaag
     1921 atttggtaga aatccggacc tttgtctctc tgattcctgt tcaaacacaa agaagaagaa
     1981 caagctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     2041 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     2101 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     2161 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     2221 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     2281 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     2341 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     2401 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     2461 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     2521 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     2581 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     2641 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     2701 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     2761 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     2821 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     2881 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     2941 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     3001 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     3061 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     3121 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     3181 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     3241 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     3301 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     3361 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     3421 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     3481 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     3541 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     3601 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     3661 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     3721 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     3781 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     3841 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3901 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3961 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4021 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     4081 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4141 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6241 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6301 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6361 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6421 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6481 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6541 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6601 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6661 aacgccaggg ttttcccagt cacgacgtt
//
