LOCUS       sequence-297006-        4796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT4G20790 P2_pECIA2 should be used with prey vector 
            AT4G20790 P2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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: AT4G20790 P2_pECIA2
FEATURES             Location/Qualifiers
     source          1..4796
                     /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             1049..1072
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1778..1819
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1829..1846
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1869..1886)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1919..2053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1943..1962)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1997..2016
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2197..2213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2197..2213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2210..2232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2221..2237
                     /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(2245..2251,2252..2269,2270..2275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2290..2311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2428..2445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2599..3187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2679..2698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3358..4149,4150..4218))
                     /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     3981..4000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4219..4323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4391..4409
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4447..4469)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4569..4588
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4782..4796,1..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 agtccacctc ctgatttcaa
      541 caccactatt tccaacaact gtcttcgtga tccaatgctc cgttactgca acaacacttc
      601 ctcttctcca atggacattg tcgagatttt cagatccaca atcgtcgcta gtcatctctg
      661 taacgagtcc aaaaacccta actgcgttga atctttccct agaatcagaa ttcacggtcg
      721 tcctaaaacc gctgctcttt acctctcttt cgatttcttc tggaagtact gtcctttaac
      781 cgtcgttgat atcgagttag tgaataactc tctaacgaat ggttttccga caaatgttct
      841 gtcttgtgct cagattcgta ctcttgatct gagttacaac caattctctg gttttgttcc
      901 ggttcaagat ctctcaggct tacccaatct gacccatttg aacctatcgt ataaccgatt
      961 ctccgagaat aagatctcgg attctgagtt cttcaaacgg tttaacgcct ctagtttcat
     1021 acagtccggt gttcttccca acgtgaagct ggaggtgctg ttccagggac cagggagcga
     1081 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     1141 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     1201 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     1261 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     1321 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     1381 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     1441 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     1501 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     1561 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     1621 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     1681 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     1741 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     1801 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     1861 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     1921 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1981 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2041 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2101 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     2161 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3001 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3961 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4321 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4381 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4441 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4501 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4561 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4621 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     4681 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     4741 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
