LOCUS       sequence-270289-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PRK4 J6_pECIA14 should be used with bait vector PRK4 
            J6_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  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 5942)
  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: PRK4 J6_pECIA14
FEATURES             Location/Qualifiers
     source          1..5942
                     /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             1238..1261
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2936..2959
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2975..2992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3015..3032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3065..3199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3089..3108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3143..3162
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3343..3359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3343..3359)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3356..3378)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3367..3383
                     /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(3391..3397,3398..3415,3416..3421))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3436..3457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3574..3591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3745..4333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3825..3844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4504..5295,5296..5364))
                     /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     5127..5146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5365..5469)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5537..5555
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5593..5615)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5715..5734
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5928..5942,1..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 tctcagccgc aggcggatgt
      541 tttgccactg ccagcctcag acgcagattg cctcttgaga tttaaagata ctttggttaa
      601 tgcatcgttc attagcagtt gggatccttc catctccccg tgtaagcgaa actcagagaa
      661 ttggttcggt gttctctgtg ttaccggtaa tgtttggggc ctacaactcg aaggaatggg
      721 cttaaccggg aagcttgacc ttgaaccatt agctgcaatc aagaatctac gaaccttgag
      781 cttcatgaac aacaaattta acggttcaat gccatctgtc aagaattttg gtgcgttgaa
      841 atcattgtac ttgtctaaca accggtttac aggggagata cccgcggatg cgtttgatgg
      901 tatgcatcat ttgaagaagc ttctgttggc taacaacgcg tttcgaggga gtatcccttc
      961 ttctttagct tatttgccaa tgcttttaga gttgaggcta aatgggaatc agtttcatgg
     1021 ggaaatacct tattttaaac aaaaggacct taagttggct agcttcgaaa acaatgacct
     1081 cgagggacct ataccggaaa gccttagcaa catggatcct gtctcctttt cagggaacaa
     1141 gaacttgtgt ggtcctccac taagcccatg ttcgagtgat tcaggatcgt ctccggatct
     1201 cccttctagt cccacggaaa agaacaagaa ccaatctctg gaggtgctgt tccagggacc
     1261 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     1321 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     1381 gaagaacacc gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc
     1441 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     1501 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     1561 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     1621 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     1681 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     1741 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     1801 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     1861 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     1921 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     1981 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     2041 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     2101 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     2161 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     2221 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     2281 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     2341 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     2401 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     2461 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     2521 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     2581 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     2641 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     2701 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     2761 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     2821 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     2881 cggcaccacc gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta
     2941 caaggatgat gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta
     3001 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     3061 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3121 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3181 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3241 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     3301 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     3361 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3421 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3481 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3541 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3721 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4381 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4441 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4501 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4621 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4801 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5341 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5461 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5521 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5581 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5641 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5701 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5761 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     5821 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5881 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5941 tt
//
