LOCUS       sequence-238696-        5948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PRK5 I7_pECIA14 should be used with bait vector PRK5 
            I7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  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 5948)
  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: PRK5 I7_pECIA14
FEATURES             Location/Qualifiers
     source          1..5948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1689..1712
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3387..3410
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3426..3443
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3466..3483)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3516..3650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3540..3559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3594..3613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3794..3810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3794..3810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3807..3829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3818..3834
                     /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(3842..3848,3849..3866,3867..3872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3887..3908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4025..4042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4196..4784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4276..4295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4955..5746,5747..5815))
                     /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     5578..5597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5816..5920)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtctca ggtggttgtg ccggattcag atgcagattg tctcttgcga ttcaaagata
     1021 cattagcaaa tggttcagaa tttcgtagtt gggatccatt atcatcacca tgtcaaggaa
     1081 atacagcaaa ttggtttggt gttctttgta gcaattacgt ttggggatta caacttgagg
     1141 gaatgggttt gaccgggaaa ctaaacctcg acccattggt tccaatgaag aatctgcgaa
     1201 ccataagctt catgaacaat aacttcaatg gcccaatgcc tcaagttaag aggtttactt
     1261 cgctgaaatc tttgtatttg tctaataacc ggttttcggg ggagataccc gcagatgcat
     1321 ttctaggtat gccacttttg aagaaaattt tgctggctaa taacgcattc cgaggaacaa
     1381 ttccttcttc tctagcctct ttgccaatgc ttttagagtt gaggctaaat ggtaatcaat
     1441 ttcaagggca aataccatct tttcaacaga aggatcttaa attagctagc tttgaaaaca
     1501 atgatctcga tggaccgata cctgaaagtc ttcgaaacat ggatccaggc tcttttgcag
     1561 gtaacaaagg tttgtgtgat gctcccttaa gtccatgttc ctcttcctct ccgggggttc
     1621 ctgttgttcc ggtatctccc gttgatccaa aatctacttc tcctcccacg gggaaaaagg
     1681 ccggatctct ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc
     1741 cccagatgct gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc
     1801 tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct
     1861 gcggcatgca gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg
     1921 acttctggaa ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac
     1981 agacagccgc caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga
     2041 cagctgccag gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg
     2101 ccatggaccg cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc
     2161 cagacagtgg agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca
     2221 ttggcttgag tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca
     2281 tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac
     2341 gagtgcagca cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact
     2401 cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc
     2461 tcatctccaa catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca
     2521 tgggaacccc agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg
     2581 ggaagaatct ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc
     2641 gcactgagct catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg
     2701 agcctggaga catgaaatac gagatccacc gagactccac actggacccc tccctgatgg
     2761 agatgacaga ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg
     2821 tggagggtgg tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg
     2881 agacgatcat gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca
     2941 cgctgagcct cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc
     3001 gagggagctc catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg
     3061 tcctcctata cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta
     3121 ccgagagcga gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag
     3181 agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc
     3241 acggcgtgca ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc
     3301 cctacaccgc ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt
     3361 ctagaggaag cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca
     3421 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     3481 ctaaggcctg agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg
     3541 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3601 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3661 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     3721 aaaacctcta caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct
     3781 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3841 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3901 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3961 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4021 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5821 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5881 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5941 cacctgac
//
