LOCUS       sequence-296429-        7244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector SKM2 X057_pECIA14 should be used with bait vector SKM2 
            X057_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7244)
  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 7244)
  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: SKM2 X057_pECIA14
FEATURES             Location/Qualifiers
     source          1..7244
                     /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             2540..2563
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4238..4261
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4277..4294
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4317..4334)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4367..4501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4391..4410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4445..4464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4645..4661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4645..4661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4658..4680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4669..4685
                     /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(4693..4699,4700..4717,4718..4723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4738..4759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4876..4893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5047..5635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5127..5146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5806..6597,6598..6666))
                     /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     6429..6448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6667..6771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6839..6857
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6895..6917)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7017..7036
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7230..7244,1..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 tccatagacg aacagggtct
      541 agctctcctc tcatggaaat ctcagttgaa catctccgga gacgctttat cttcttggaa
      601 agcctccgaa tcgaatcctt gtcaatgggt gggaatcaaa tgcaacgaaa gaggtcaagt
      661 ctcggagata caactccaag tcatggattt tcaaggtccg ttaccggcga cgaatctccg
      721 acagataaag tctctgactt tgctcagctt aacctccgtg aatctcactg gttcaatccc
      781 taaggagctt ggagacttgt cggagctaga agttcttgat ttagctgata attctctctc
      841 tggtgaaatc cctgtggata tcttcaagct caagaaacta aagattctgt ctctcaacac
      901 aaataatctc gaaggtgtga ttccatcgga gttagggaat ctcgttaacc tcatcgagct
      961 cactctcttc gacaataaac tcgccggaga gattcctaga accatcggag aactcaagaa
     1021 tctagaaatc ttccgtgcag gtggtaacaa gaatctcaga ggagagcttc cttgggagat
     1081 tggtaactgc gagagtcttg tgactttagg tctcgccgaa acaagtctct ccggtagact
     1141 tccggcgtcg atcggaaacc tgaaaaaggt tcagacaata gctctttaca cgtcactctt
     1201 gtctggtcca atccctgatg agattggaaa ctgcacagag cttcaaaacc tctacttgta
     1261 ccaaaactca atctctggat cgattcctgt atcaatggga cgtctcaaga agcttcaaag
     1321 tcttctctta tggcaaaaca atctcgtcgg aaagatccct accgagctag gcacctgtcc
     1381 ggagcttttc ctcgtcgatt tatcggaaaa tctcctcacc ggaaacatcc caagaagctt
     1441 cggaaacctt ccaaatctcc aagagcttca gctaagtgtt aaccagctct ccggtacaat
     1501 ccctgaagaa ctcgcgaatt gcacgaagtt gacgcattta gagatcgata acaaccaaat
     1561 ctccggcgag attccgccgt tgatcgggaa attaacaagc ttgacgatgt tctttgcgtg
     1621 gcagaatcag ttaaccggaa taattccaga gtctctttct caatgtcaag agcttcaagc
     1681 gattgatctc tcctacaaca atctctctgg ttcgatacct aatggaatct ttgagattcg
     1741 aaacctcacg aagcttcttc ttctctctaa ctacttatct ggattcatac caccggatat
     1801 tggaaactgc acgaatcttt accggttaag actcaacggt aacagactcg cggggaatat
     1861 tccggcggag attgggaatc tgaaaaatct caacttcatc gatataagtg agaatcgtct
     1921 catcggaaac attcctccgg agatctccgg ttgtacaagt ctcgaattcg tcgatctaca
     1981 ttcaaacggt ttaaccggtg gtttacccgg tacacttccg aagagcttac aattcattga
     2041 tttatccgat aattcattaa ccggttctct accaaccgga atcggatcgt taaccgaact
     2101 cacgaagctc aatctcgcga agaatcgttt ctccggcgaa atccctcgag agatatcttc
     2161 ctgtcggagt cttcagcttc tcaacctcgg cgacaatggc ttcaccggag aaatccctaa
     2221 cgagttagga cggatcccgt ctcttgcgat ctctctaaat ctcagctgca atcacttcac
     2281 cggagagatc ccgtcgagat tctctagcct cactaatctc ggaaccctcg atgtctctca
     2341 caacaaactc gccggaaacc taaacgtctt agctgatcta cagaacctcg tctcactcaa
     2401 catctccttc aacgaattct caggcgagtt accaaatact ctgtttttca ggaaacttcc
     2461 actctctgtt cttgaatcaa acaaagggct atttatttcg acccgacccg agaatgggat
     2521 ccaaacccga caccgttcgc tggaggtgct gttccaggga ccaagcggca atcccggcgg
     2581 cgatctggcc ccccagatgc tgcgcgagct gcaggagacc aatgccgccc tgcaggacgt
     2641 gcgcgagctg ctgcgccacc gcgtgaagga gatcaccttc ctgaagaaca ccgtgatgga
     2701 gtgcgacgcc tgcggcatgc agccctctag aggctccggc tccatcatcc cagttgagga
     2761 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     2821 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     2881 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     2941 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     3001 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     3061 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     3121 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     3181 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     3241 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     3301 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtggag gccgaaagta
     3361 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     3421 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     3481 tgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     3541 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     3601 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     3661 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     3721 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     3781 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     3841 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     3901 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     3961 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     4021 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     4081 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     4141 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     4201 gcacccgggt tctagaggaa gcacctccgg cgataacgat tacaaggatg atgatgataa
     4261 ggccagcggc accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     4321 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     4381 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4441 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4501 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4561 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     4621 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4681 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4741 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4801 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4861 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5041 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5101 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5161 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5221 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5281 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5341 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5401 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5461 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5521 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5581 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5641 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5701 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5761 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5941 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6661 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6781 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6841 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6901 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6961 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     7021 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     7081 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     7141 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     7201 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
