LOCUS       sequence-296840-        5198 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector AT4G34220 X018_pECIA2 should be used with prey vector 
            AT4G34220 X018_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5198)
  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 5198)
  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: AT4G34220 X018_pECIA2
FEATURES             Location/Qualifiers
     source          1..5198
                     /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             1451..1474
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2180..2221
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2231..2248
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2271..2288)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2321..2455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2345..2364)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2399..2418
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2599..2615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2599..2615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2612..2634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2623..2639
                     /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(2647..2653,2654..2671,2672..2677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2692..2713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2830..2847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3001..3589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3081..3100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3760..4551,4552..4620))
                     /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     4383..4402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4621..4725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4793..4811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4849..4871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4971..4990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5184..5198,1..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 aacactgatg gtgttcttct
      541 gcttactttc aaatactcca tcctcactga ccctctctct gttctacgca attggaacta
      601 tgatgatgca acaccatgct tgtggacagg tgtcacatgt acagagcttg ggaagcctaa
      661 cacaccagac atgttcagag tcacaagctt ggtacttccc aacaaacacc ttttgggttc
      721 catcactccg gacttgttct cgattccata tctaaggatc ttggatctgt ctagcaattt
      781 cttcaacggg tcactcccgg actcggtgtt caacgccact gagcttcaga gtatctccct
      841 tggaagcaac aatctctctg gtgatttgcc caagagtgtc aacagtgtga ctaatctcca
      901 gctcttgaat ctctcagcta acgcatttac tggcgaaatt ccgctcaaca tctcacttct
      961 gaagaatcta acggtcgttt ccttgtcaaa aaacaccttt tcaggtgata ttccaagtgg
     1021 tttcgaagca gcacagattc ttgatctttc ttcaaatctt ctcaacggct ctcttcccaa
     1081 ggatttggga ggaaaaagcc tgcattactt gaacttatca cacaacaaag tcctcggcga
     1141 gatttctcca aattttgcag agaagtttcc agcaaatgcc acagtcgatc tctcctttaa
     1201 caacctcaca ggcccaatcc caagttccct ttctttactt aaccaaaagg ctgagtcttt
     1261 ttccggtaac caagagcttt gtgggaagcc attgaagatc ctttgttcaa ttccttccac
     1321 tctttcaaac cctccaaata tctctgaaac tacttcacca gcaatagcag ttaaaccaag
     1381 aagtacagct ccaataaacc ctttgacaga aaaaccaaat cagacaggaa aaagcaagct
     1441 aaagccaagc ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     1501 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     1561 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     1621 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     1681 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     1741 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     1801 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     1861 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     1921 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     1981 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2041 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     2101 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     2161 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2221 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2281 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     2341 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2401 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2461 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2521 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     2581 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5041 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5101 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5161 aagttgggta acgccagggt tttcccagtc acgacgtt
//
