LOCUS       sequence-298803-        6392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G17230 X074_pECIA2 should be used with prey vector 
            AT1G17230 X074_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6392)
  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 6392)
  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: AT1G17230 X074_pECIA2
FEATURES             Location/Qualifiers
     source          1..6392
                     /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             2645..2668
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3374..3415
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3425..3442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3465..3482)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3515..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3539..3558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3593..3612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3793..3809)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3793..3809)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3806..3828)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3817..3833
                     /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(3841..3847,3848..3865,3866..3871))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3886..3907
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4024..4041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4195..4783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4275..4294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4954..5745,5746..5814))
                     /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     5577..5596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5815..5919)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5987..6005
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6043..6065)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6165..6184
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6378..6392,1..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 tccttaaacg aagaagggcg
      541 tgttcttctg gagtttaaag ctttccttaa tgactccaat ggctatctcg caagctggaa
      601 tcagttggat tcaaatccct gcaactggac cggaatcgcg tgtacccatc tcagaactgt
      661 aacttctgtt gatctaaatg ggatgaatct atcaggtaca ctatctcctc tcatatgcaa
      721 gcttcatggt ttaaggaaat tgaatgtctc caccaacttc atctctggtc caatccctca
      781 agatttatct ctctgtcgaa gtctagaggt tcttgatctc tgcacaaata ggttccatgg
      841 agtgatacca attcagctaa ccatgatcat cactcttaag aagctgtatc tatgcgagaa
      901 ttatcttttc ggttctatcc ctagacaaat cggtaatttg agttcactcc aagagcttgt
      961 gatctacagc aataatctca ccggtgtaat ccctccttcc atggcgaaat tgagacagct
     1021 ccggatcatt agggcaggac ggaatggatt ctcgggtgtg attccgtctg aaatcagcgg
     1081 atgcgagagc ctaaaggtac ttggattagc agagaatctt cttgagggtt ctcttcctaa
     1141 gcagctcgag aagcttcaaa atcttactga tttgatactc tggcaaaacc gtctatctgg
     1201 tgagatacct ccttccgtag gaaacatcag cagactggag gtgcttgcgc tgcatgaaaa
     1261 ctacttcacg ggatcaattc caagagagat tgggaagctc acaaagatga aaaggctgta
     1321 cctttacaca aaccagttaa ccggagagat acctcgcgaa ataggcaact tgatagatgc
     1381 agctgagata gatttttcgg agaatcagtt aacaggtttc ataccaaaag agtttggtca
     1441 tatcctgaat ctcaaattac ttcatctttt tgagaacatt cttctaggtc caattcctag
     1501 ggagcttggg gaattgacac tgttggagaa gctagacttg tctatcaata gactaaacgg
     1561 tactatccca caagagcttc agttcctacc ttaccttgta gatttgcagc tttttgacaa
     1621 ccagcttgaa gggaagatac ctcctttaat cggtttctat agcaattttt ctgttcttga
     1681 catgtctgcc aattctctgt ctggtccaat tcctgcccac ttctgcagat ttcagacgct
     1741 aatacttctg agtcttggtt caaacaagtt atcaggaaac atccctcgtg atctgaaaac
     1801 ctgcaagtca ctgacaaagc taatgctagg cgataaccag ctaacaggaa gccttcccat
     1861 tgaattattt aatcttcaga accttactgc attagagctt caccaaaact ggttatcagg
     1921 gaatatatct gcagacctgg gaaagctgaa gaatttggaa agactgcgtc tagctaacaa
     1981 caattttact ggtgaaattc ctcctgagat tgggaatctc acaaagattg ttggctttaa
     2041 catctcctcc aatcagctca ctggtcacat ccctaaagag ttgggaagtt gtgtcactat
     2101 ccagaggctt gaccttagtg gcaacaagtt ttccggatac atagcacaag agcttggtca
     2161 gttggtatat ttggagattt tgagattgtc tgataataga ttgaccggag aaattcctca
     2221 tagctttgga gatcttactc ggctcatgga gttacaacta ggaggcaatc tcttgtctga
     2281 aaacatccca gtggagctcg ggaagttaac ctctctgcaa atctccctca acattagtca
     2341 caataatctc tcaggtacta ttccagacag tttaggaaat ctacagatgt tggagatact
     2401 ttacctcaac gacaacaagc tttcaggtga gatccctgca tcaattggga atctcatgag
     2461 cctcctcatc tgcaatatct caaacaataa cttggtcgga acagtaccag acacagccgt
     2521 gttccagcgg atggattcat cgaactttgc tggaaaccat ggtttgtgta actctcagag
     2581 aagccattgt caaccacttg tcccacattc tgattcaaag ctaaattggt taatcaacgg
     2641 ttcgctggag gtgctgttcc agggaccagg gagcgacaaa actcacacat gcccaccgtg
     2701 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     2761 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     2821 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     2881 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     2941 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     3001 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     3061 caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt
     3121 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     3181 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     3241 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     3301 tgagggtctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatctag
     3361 agggcccttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     3421 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3481 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3541 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3601 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3661 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3721 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     3781 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3841 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3901 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3961 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4021 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4081 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4141 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4201 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4261 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4321 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4381 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4441 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4501 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4561 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4621 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4681 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4741 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4801 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4861 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4921 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4981 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5041 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5101 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5161 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5221 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5281 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5341 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5401 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5461 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5521 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5581 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5641 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5701 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5761 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5881 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5941 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6001 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6061 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6121 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6181 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6241 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6301 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6361 ggtaacgcca gggttttccc agtcacgacg tt
//
