LOCUS       sequence-269949-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G67720 M7_pECIA14 should be used with bait vector 
            AT1G67720 M7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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 6746)
  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: AT1G67720 M7_pECIA14
FEATURES             Location/Qualifiers
     source          1..6746
                     /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             2042..2065
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3740..3763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3779..3796
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3819..3836)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3869..4003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3893..3912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3947..3966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4147..4163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4147..4163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4160..4182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4171..4187
                     /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(4195..4201,4202..4219,4220..4225))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4240..4261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4378..4395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4549..5137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4629..4648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5308..6099,6100..6168))
                     /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     5931..5950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6169..6273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6341..6359
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6397..6419)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6519..6538
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6732..6746,1..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 tctcaagtta cagagtttgt
      541 aagtatagat tgtggttgct caagtaacta cacagaccca agaactggac taggatgggt
      601 ttcagattca gagatcatca aacagggcaa gccagtaact ttggccaata ccaattggaa
      661 ctcgatgcaa taccgtagga gaagagattt cccaactgac aataagaagt attgttacag
      721 actcagtacc aaagagagaa ggcgatacat tgtcagaact acatttcttt atggtggttt
      781 gggtagtgag gaagcctacc caaagtttca actttacttg gatgcaacga aatgggctac
      841 tgtgacgatt caggaggtct ctagggttta tgttgaggag ttgatcgtaa gggcaacttc
      901 aagttatgtg gatgtttgtg tatgctgtgc tatcaccggt tctcctttca tgtccactct
      961 tgagcttcgc ccgttgaacc tttcgatgta tgccactgac tacgaggata atttcttctt
     1021 gaaggttgct gccagagtga attttggtgc tccgaacatg gatgccttga ggtacccaga
     1081 tgatccatat gacagaatct gggagtcaga tattaacaaa cggccaaact atcttgttgg
     1141 tgtggctcct ggaacgacaa gaatcaatac gtcaaagact ataaatacgt tgactagaga
     1201 gtatccgcct atgaaagtca tgcaaacagc agtagttggc acacaaggac tgatcagcta
     1261 tagattgaac ctggaagact tccctgccaa tgctcgtgcg tatgcttatt ttgctgaaat
     1321 tgaagaactt ggtgctaacg agacccggaa atttaagttg gtgcagccat actttcctga
     1381 ctatagtaat gcagtggtga acattgccga gaatgccaat gggagctata ctctctatga
     1441 acctagctat atgaatgtga ctttggattt tgttttgacg ttctcatttg ggaagactaa
     1501 ggattctacg caagggccac tcctaaatgc aattgaaatt agcaagtatc taccaatatc
     1561 cgtgaaaact gataggagtg acgtgtctgt tcttgatgca attcgttcga tgtcccctga
     1621 tagcgattgg gccagtgaag gaggagaccc ttgtatccca gttctttggt cgtgggtaaa
     1681 ctgtagctca acttcaccac caagagtcac gaaaattgct ctatcaagga aaaatctgag
     1741 gggtgagatt ccgcctggga tcaattatat ggaggcattg acagagttgt ggcttgacga
     1801 taatgaattg accggaactc ttccagacat gagcaaactt gtcaacctga aaatcatgca
     1861 tctagaaaac aaccaactta gtggttcact cccaccatac cttgctcatc tgcctaacct
     1921 acaagaactg tctatagaaa ataactcctt taaaggaaaa attccttcag cgctattaaa
     1981 ggggaaagtt ttgttcaaat ataataataa tcctgagctt cagaacgagg cacagcgaaa
     2041 gctggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg ccccccagat
     2101 gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca
     2161 ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg cctgcggcat
     2221 gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc cggacttctg
     2281 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     2341 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     2401 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     2461 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     2521 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     2581 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     2641 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     2701 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     2761 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     2821 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     2881 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     2941 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     3001 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     3061 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     3121 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     3181 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     3241 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     3301 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     3361 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     3421 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     3481 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     3541 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     3601 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     3661 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gttctagagg
     3721 aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg gcaccggtca
     3781 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3841 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3901 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3961 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4021 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4081 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     4141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6301 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6361 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6421 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6481 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6541 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6601 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6661 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6721 gccagggttt tcccagtcac gacgtt
//
