LOCUS       sequence-276503-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector CLV1 L2_pECIA14 should be used with bait vector CLV1 
            L2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  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 7079)
  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: CLV1 L2_pECIA14
FEATURES             Location/Qualifiers
     source          1..7079
                     /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             2375..2398
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4073..4096
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4112..4129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4152..4169)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4202..4336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4226..4245)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4280..4299
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4480..4496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4480..4496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4493..4515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4504..4520
                     /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(4528..4534,4535..4552,4553..4558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4711..4728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4882..5470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4962..4981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5641..6432,6433..6501))
                     /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     6264..6283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6502..6606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6674..6692
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6730..6752)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6852..6871
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7065..7079,1..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 tcaccatgtt tcgcttacac
      541 tgacatggaa gttcttctca atctcaaatc ctccatgatt ggtcctaaag gacacggtct
      601 ccacgactgg attcactcat cttctccgga tgctcactgt tctttctccg gcgtctcatg
      661 tgacgacgat gctcgtgtta tctctctcaa cgtctccttc actcctttgt ttggtacaat
      721 ctcaccagag attgggatgt tgactcattt ggtgaatcta actttagctg ccaacaactt
      781 caccggtgaa ttaccattgg agatgaagag tctaacttct ctcaaggttt tgaatatctc
      841 caacaatggt aaccttactg gaacattccc tggagagatt ttaaaagcta tggttgatct
      901 tgaagttctt gacacttata acaacaattt caacggtaag ttaccaccgg agatgtcaga
      961 gcttaagaag cttaaatacc tctctttcgg tggaaatttc ttcagcggag agattccaga
     1021 gagttatgga gatattcaaa gcttagagta tcttggtctc aacggagctg gactctccgg
     1081 taaatctccg gcgtttcttt cccgcctcaa gaacttaaga gaaatgtata ttggctacta
     1141 caacagctac accggtggtg ttccaccgga gttcggtggt ttaacaaagc ttgagatcct
     1201 cgacatggcg agctgtacac tcaccggaga gattccgacg agtttaagta acctgaaaca
     1261 tctacatact ctgtttcttc acatcaacaa cttaaccggt catataccac cggagctttc
     1321 cggtttagtc agcttgaaat ctctcgattt atcaatcaat cagttaaccg gagaaatccc
     1381 tcaaagcttc atcaatctcg gaaacattac tctaatcaat ctcttcagaa acaatctcta
     1441 cggacaaata ccagaggcca tcggagaatt accaaaactc gaagtcttcg aagtatggga
     1501 gaacaatttc acgttacaat taccggcgaa tcttggccgg aacgggaatc taataaagct
     1561 tgatgtctct gataatcatc tcaccggact tatccccaag gacttatgca gaggtgagaa
     1621 attagagatg ttaattctct ctaacaactt cttctttggt ccaattccag aagagcttgg
     1681 taaatgcaaa tccttaacca aaatcagaat cgttaagaat cttctcaacg gcactgttcc
     1741 ggcggggctt ttcaatctac cgttagttac gattatcgaa ctcactgata atttcttctc
     1801 cggtgaactt ccggtaacga tgtccggcga tgttctcgat cagatttacc tctctaacaa
     1861 ctggttttcc ggcgagattc cacctgcgat tggtaatttc cccaatctac agactctatt
     1921 cttagatcgg aaccgatttc gcggcaacat tccgagagaa atcttcgaat tgaagcattt
     1981 atcgaggatc aacacaagtg cgaacaacat caccggcggt attccagatt caatctctcg
     2041 ctgctcaact ttaatctccg tcgatctcag ccgtaaccga atcaacggag aaatccctaa
     2101 agggatcaac aacgtgaaaa acttaggaac tctaaatatc tccggtaatc aattaaccgg
     2161 ttcaatccct accggaatcg gaaacatgac gagtttaaca actctcgatc tctctttcaa
     2221 cgatctctcc ggtagagtac cactcggtgg tcaattcttg gtgttcaacg aaacttcctt
     2281 cgccggaaac acttacctct gtctccctca ccgtgtctct tgtccaacac ggccaggaca
     2341 aacctccgat cacaatcaca cggcgttgtt ctcactggag gtgctgttcc agggaccaag
     2401 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc
     2461 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa
     2521 gaacaccgtg atggagtgcg acgcctgcgg catgcagccc tctagaggct ccggctccat
     2581 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     2641 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     2701 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     2761 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     2821 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     2881 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     2941 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     3001 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     3061 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     3121 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     3181 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     3241 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     3301 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     3361 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     3421 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     3481 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     3541 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     3601 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     3661 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     3721 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     3781 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     3841 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     3901 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     3961 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     4021 caccaccgac gccgcgcacc cgggttctag aggaagcacc tccggcgata acgattacaa
     4081 ggatgatgat gataaggcca gcggcaccgg tcatcatcac catcaccatt gagtttaaac
     4141 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     4201 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4261 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4321 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4381 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     4441 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6601 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6661 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     6721 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     6781 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6841 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     6901 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     6961 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7021 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
