LOCUS       sequence-296816-        4964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector PRK6 K3_pECIA2 should be used with prey vector PRK6 
            K3_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4964)
  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 4964)
  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: PRK6 K3_pECIA2
FEATURES             Location/Qualifiers
     source          1..4964
                     /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             1217..1240
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1946..1987
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1997..2014
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2037..2054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2087..2221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2111..2130)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2165..2184
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2365..2381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2365..2381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2378..2400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2389..2405
                     /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(2413..2419,2420..2437,2438..2443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2458..2479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2596..2613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2767..3355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2847..2866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3526..4317,4318..4386))
                     /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     4149..4168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4387..4491)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4559..4577
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4615..4637)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4737..4756
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4950..4964,1..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 tccgaaccgc ttgtgcggtt
      541 caaaaactcg gtgaagataa ccaaagggga cctaaattca tggagagaag ggacagatcc
      601 ttgcagtggg aaatggtttg ggatctattg ccaaaagggt ctaacagtct caggcattca
      661 cgtcacacgt ctaggtctct caggaaccat caccgtggat gatttaaagg atctcccaaa
      721 tctaaagacg atcaggcttg acaacaacct cctctccggt cctctcccgc atttctttaa
      781 actccgtggc ctaaaatctc tcatgctctc taacaacagc ttctccggag agatccgtga
      841 cgattttttc aaggacatgt caaagctcaa gcgactattt cttgatcata acaagttcga
      901 gggaagtatc ccttcctcca ttacgcagct ccctcaactc gaggagcttc acatgcaaag
      961 taataacctc accggagaga tacctccaga gtttggcagc atgaagaacc ttaaagtcct
     1021 cgacctctca accaacagcc tcgatggaat cgtcccgcaa agcattgcgg ataagaaaaa
     1081 tctcgctgtt aatttgacag agaacgaata cttgtgcggg ccggtggtgg acgttggatg
     1141 tgagaatatt gagctaaacg atccgcaaga aggacaaccc ccgtcgaaac catcctcctc
     1201 ggtaccagaa acctcgctgg aggtgctgtt ccagggacca gggagcgaca aaactcacac
     1261 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     1321 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     1381 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     1441 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     1501 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     1561 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     1621 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     1681 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     1741 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     1801 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     1861 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     1921 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     1981 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     2041 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     2101 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2161 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2221 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2281 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     2341 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4501 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4561 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4621 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4681 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4741 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4801 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4861 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4921 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
