LOCUS       sequence-296770-        4973 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector RKL1 I6_pECIA2 should be used with prey vector RKL1 
            I6_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4973)
  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 4973)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_pECIA2
FEATURES             Location/Qualifiers
     source          1..4973
                     /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             1955..1996
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2006..2023
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2046..2063)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2096..2230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2120..2139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2174..2193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2374..2390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2374..2390)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2387..2409)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2398..2414
                     /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(2422..2428,2429..2446,2447..2452))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2467..2488
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2605..2622)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2776..3364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2856..2875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3535..4326,4327..4395))
                     /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     4158..4177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4396..4500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4568..4586
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4624..4646)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4746..4765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4959..4973,1..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 cagggaccag ggagcgacaa
     1261 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     1321 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     1381 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     1441 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     1501 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     1561 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     1621 gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga ccaagaacca
     1681 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     1741 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     1801 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     1861 cttctcatgc tccgtgatgc atgagggtct gcacaaccac tacacgcaga agagcctctc
     1921 cctgtctccg ggtaaatcta gagggccctt cgaaggtaag cctatcccta accctctcct
     1981 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     2041 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     2101 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2161 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2221 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2281 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     2341 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2401 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2461 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2521 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2941 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3181 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3601 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4501 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4561 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4621 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4681 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4741 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4801 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     4861 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4921 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
