LOCUS       sequence-269972-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RKL1 I6_pECIA14 should be used with bait vector RKL1 
            I6_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  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 5930)
  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: RKL1 I6_pECIA14
FEATURES             Location/Qualifiers
     source          1..5930
                     /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             1226..1249
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2924..2947
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2963..2980
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3003..3020)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3053..3187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3077..3096)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3131..3150
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3331..3347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3344..3366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3355..3371
                     /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(3379..3385,3386..3403,3404..3409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3424..3445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3562..3579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4492..5283,5284..5352))
                     /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     5115..5134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5353..5457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5525..5543
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5581..5603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5703..5722
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5916..5930,1..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 caagatctca acgccgacag
      541 aaccgctctt ctctctctcc gctccgccgt cggaggccgt acatttcgct ggaacatcaa
      601 acaaacctcg ccgtgtaact gggccggtgt caaatgcgag agtaacagag tcaccgctct
      661 ccgcttaccc ggcgttgctc tctccggtga cattccagag ggtattttcg gtaacttaac
      721 tcagctccgt acacttagcc ttcgtctcaa tgctctctct ggctcactcc ctaaagatct
      781 cagtacctct tccaatctcc gacaccttta cttacaagga aacagattct ccggcgagat
      841 acctgaagtt ttgttcagtc tcagtcacct cgtgaggctt aatctcgctt ccaacagctt
      901 caccggagaa atctcttcag gtttcacgaa cctcacgaag ctaaagactc tgtttctaga
      961 gaataatcaa ctctctggtt caatccctga cttagattta cctttggttc aattcaacgt
     1021 ttcgaataac tcgttgaacg gttctatacc taagaatctt cagaggtttg agtctgattc
     1081 gtttctacag acttcacttt gtggcaaacc gcttaaactt tgtcctgatg aagaaactgt
     1141 acctagtcag cctacttctg gtggaaacag aacaccaccg agtgttgaag gaagtgaaga
     1201 gaagaagaaa aagaacaaac tttccctgga ggtgctgttc cagggaccaa gcggcaatcc
     1261 cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg ccgccctgca
     1321 ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga agaacaccgt
     1381 gatggagtgc gacgcctgcg gcatgcagcc ctctagaggc tccggctcca tcatcccagt
     1441 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     1501 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     1561 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     1621 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     1681 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     1741 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     1801 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     1861 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     1921 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     1981 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     2041 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     2101 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     2161 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     2221 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     2281 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     2341 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     2401 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     2461 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     2521 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     2581 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     2641 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     2701 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     2761 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     2821 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     2881 cgccgcgcac ccgggttcta gaggaagcac ctccggcgat aacgattaca aggatgatga
     2941 tgataaggcc agcggcaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     3001 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3061 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3121 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3181 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3241 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3301 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5461 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5521 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5581 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5641 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5701 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5761 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5821 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5881 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
