LOCUS       sequence-296844-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector SKM1 nA12_pECIA2 should be used with prey vector SKM1 
            nA12_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  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 6074)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6074
                     /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             3056..3097
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3107..3124
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3147..3164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3197..3331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3221..3240)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3275..3294
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3475..3491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3475..3491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3488..3510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3499..3515
                     /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(3523..3529,3530..3547,3548..3553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3568..3589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3706..3723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3957..3976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4636..5427,5428..5496))
                     /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     5259..5278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5497..5601)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5669..5687
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5725..5747)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5847..5866
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6060..6074,1..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 gggagcgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct
     2401 ggggggaccg tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg
     2461 gacccctgag gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt
     2521 caactggtac gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca
     2581 gtacaacagc acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa
     2641 tggcaaggag tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac
     2701 catctccaaa gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg
     2761 ggaggagatg accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag
     2821 cgacatcgcc gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc
     2881 tcccgtgctg gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag
     2941 caggtggcag caggggaacg tcttctcatg ctccgtgatg catgagggtc tgcacaacca
     3001 ctacacgcag aagagcctct ccctgtctcc gggtaaatct agagggccct tcgaaggtaa
     3061 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     3121 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     3181 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3241 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3301 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3361 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3421 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     3481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5641 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5701 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5761 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5821 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5881 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5941 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6001 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6061 ccagtcacga cgtt
//
