LOCUS       sequence-270518-        7031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector SKM1 nA12_pECIA14 should be used with bait vector SKM1 
            nA12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7031)
  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 7031)
  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: SKM1 nA12_pECIA14
FEATURES             Location/Qualifiers
     source          1..7031
                     /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             2327..2350
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4025..4048
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4064..4081
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4104..4121)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4154..4288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4232..4251
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4432..4448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4432..4448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4445..4467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4456..4472
                     /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(4480..4486,4487..4504,4505..4510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4525..4546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4663..4680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4834..5422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4914..4933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5593..6384,6385..6453))
                     /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     6216..6235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6454..6558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6626..6644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6682..6704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6804..6823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7017..7031,1..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 catgcaaatg aactcgagct
      541 tctcttatca ttcaaatcct caattcaaga ccctctaaaa cacctctcct cttggtctta
      601 ctcttcaacc aatgatgtgt gtctttggag tggcgtcgtt tgcaacaaca tctctcgtgt
      661 tgtctctctc gatctctccg gcaagaacat gtccggtcaa atcctcaccg ccgctacttt
      721 tcgcctacct tttcttcaaa ctatcaacct ctcaaacaac aacttgtccg gtccaatacc
      781 tcacgatatc tttaccactt cgtctccttc tctccgttac ctcaacttaa gcaacaacaa
      841 cttttccggc tccatccctc gaggctttct tccgaacctc tacacgttag atctttccaa
      901 caacatgttc accggtgaaa tctacaacga catcggggtt ttctctaacc ttagagtcct
      961 tgatctcggt ggaaatgtct taaccggcca tgtaccgggc tatcttggaa acctctcaag
     1021 attagagttc ttaacattgg cttcaaacca gttaaccgga ggtgtaccgg tggaattagg
     1081 gaagatgaaa aacttgaaat ggatatactt gggatacaac aatctctcag gagaaatccc
     1141 ttaccaaatc ggtgggttat cctctcttaa ccaccttgat ctcgtctaca acaatctctc
     1201 tggaccgata cctccttcac taggtgactt gaagaagctt gagtatatgt ttctctatca
     1261 aaacaagctc tccggtcaga tcccaccgtc catcttctcc ctccaaaacc taatttctct
     1321 cgactttagc gataactcgc tctcgggaga gatccctgag ctcgtggctc agatgcaaag
     1381 cttggagatt ctccatctct tctctaataa cttaacagga aaaattccag aaggagtaac
     1441 ttctttgcca cgtctaaaag ttcttcagct ttggtcaaac agattttccg gtggaattcc
     1501 ggcaaatctt gggaaacata acaatctcac tgttcttgat ctttctacca acaatctcac
     1561 cggaaaactc cctgacactc tttgtgattc tggccatctc actaagctca tcctcttctc
     1621 taactctcta gacagccaaa tcccaccgag tttgggcatg tgtcagagct tagaacgcgt
     1681 gaggcttcaa aacaatggct tttcaggcaa gttacctcga ggtttcacca agttgcaact
     1741 cgttaacttt ttggacttat ccaacaacaa cctccaaggt aacatcaaca catgggacat
     1801 gccacaactt gagatgttag acctcagtgt gaacaaattc ttcggtgaat taccggattt
     1861 ttctagaagc aaaagactca agaagctcga tttatcacgg aacaaaatct ctggagtggt
     1921 tccacaaggg ctcatgacgt ttccggaaat aatggatttg gatttaagtg agaacgaaat
     1981 cactggagtg atcccaaggg agttgtcttc ttgcaaaaat ctagtgaatc ttgacttgag
     2041 ccacaacaat tttaccggag agatcccttc gagtttcgca gaattccaag ttctcagcga
     2101 tcttgatttg tcatgcaacc aattatcagg cgagattccg aagaatctag gaaacattga
     2161 gtctcttgtt caagtcaaca tctctcataa tcttcttcac ggaagcttac ctttcaccgg
     2221 agcttttctt gccataaacg caacagcagt tgagggtaac atcgatcttt gtagcgaaaa
     2281 ttccgctagc ggtttacgtc cttgcaaggt tgtaagaaag agaagtctgg aggtgctgtt
     2341 ccagggacca agcggcaatc ccggcggcga tctggccccc cagatgctgc gcgagctgca
     2401 ggagaccaat gccgccctgc aggacgtgcg cgagctgctg cgccaccgcg tgaaggagat
     2461 caccttcctg aagaacaccg tgatggagtg cgacgcctgc ggcatgcagc cctctagagg
     2521 ctccggctcc atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     2581 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     2641 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     2701 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     2761 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     2821 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     2881 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     2941 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     3001 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     3061 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     3121 cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     3181 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     3241 gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc
     3301 cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga
     3361 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     3421 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     3481 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     3541 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     3601 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     3661 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     3721 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     3781 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     3841 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     3901 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     3961 gccccccgcc ggcaccaccg acgccgcgca cccgggttct agaggaagca cctccggcga
     4021 taacgattac aaggatgatg atgataaggc cagcggcacc ggtcatcatc accatcacca
     4081 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     4141 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     4201 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4261 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4321 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     4381 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     4441 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4501 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4561 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4621 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4681 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4741 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5521 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5581 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5641 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5701 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5761 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5821 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5881 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5941 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6001 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6061 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6121 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6181 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6241 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6301 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6361 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6421 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6481 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6541 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6601 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6661 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6721 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6781 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6841 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6901 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6961 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7021 gtcacgacgt t
//
