LOCUS       sequence-298812-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector CLV1 L2_pECIA2 should be used with prey vector CLV1 
            L2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  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 6122)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6122
                     /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             3104..3145
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3155..3172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3195..3212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3245..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3323..3342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3523..3539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3523..3539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3536..3558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3547..3563
                     /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(3571..3577,3578..3595,3596..3601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3616..3637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3754..3771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3925..4513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4005..4024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4684..5475,5476..5544))
                     /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     5307..5326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5545..5649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5717..5735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5773..5795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5895..5914
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6108..6122,1..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 agggaccagg
     2401 gagcgacaaa actcacacat gcccaccgtg cccagcacct gaactcctgg ggggaccgtc
     2461 agtcttcctc ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt
     2521 cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt
     2581 ggacggcgtg gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac
     2641 gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta
     2701 caagtgcaag gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc
     2761 caaagggcag ccccgagaac cacaggtgta caccctgccc ccatcccggg aggagatgac
     2821 caagaaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt
     2881 ggagtgggag agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga
     2941 ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca
     3001 ggggaacgtc ttctcatgct ccgtgatgca tgagggtctg cacaaccact acacgcagaa
     3061 gagcctctcc ctgtctccgg gtaaatctag agggcccttc gaaggtaagc ctatccctaa
     3121 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
     3181 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     3241 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3301 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3361 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3421 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     3481 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     3541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3781 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4501 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4621 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5701 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5761 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5821 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5881 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5941 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6001 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6061 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6121 tt
//
