LOCUS       sequence-274075-        7412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G75640 X061_pECIA14 should be used with bait vector 
            AT1G75640 X061_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  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 7412)
  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: AT1G75640 X061_pECIA14
FEATURES             Location/Qualifiers
     source          1..7412
                     /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             2708..2731
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4406..4429
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4445..4462
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4485..4502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4535..4669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4559..4578)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4613..4632
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4813..4829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4813..4829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4826..4848)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4837..4853
                     /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(4861..4867,4868..4885,4886..4891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4906..4927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5044..5061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5215..5803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5295..5314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5974..6765,6766..6834))
                     /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     6597..6616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6835..6939)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7007..7025
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7063..7085)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7185..7204
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7398..7412,1..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 tcatcagaga ctcaagcact
      541 cacatctttc aaactctctc tccacgaccc tctaggagct ctcgagtcct ggaatcaatc
      601 atctccatca gctccatgcg actggcacgg cgtttcttgc ttctccggcc gtgtccgtga
      661 gcttcgtctt cctcgtctcc acctcaccgg ccacctcagt cctcgactcg gtgagttgac
      721 tcagctccgt aaactaagtc ttcacaccaa cgacataaac ggcgccgttc cttcgtctct
      781 ctctcgttgt gtcttcctta gagctcttta tcttcactac aactccttct ccggcgattt
      841 tccaccggag atactgaatc tacgaaacct ccaggtcctc aacgctgctc acaactcact
      901 caccggaaat ctatccgacg tcacagtctc gaagagcctt cgttacgtcg atttgtcttc
      961 caatgcaata tccggtaaga ttccggcgaa tttctcagcg gattcatctc tccagctcat
     1021 caacctctcg ttcaaccact tctccggtga gatcccggct acattgggac aactacaaga
     1081 cctcgagtat ctctggctag actcaaacca attacaaggc acaataccat ccgcattagc
     1141 caactgttca tcgcttatac acttcagcgt aaccggtaat cacctcaccg gtttaattcc
     1201 ggttacttta ggcacgattc gaagcctgca ggtgatttca ttatcggaga attcatttac
     1261 cggtacagtt ccggttagtc ttctatgtgg ttattcaggt tacaattcct ccatgcgaat
     1321 cattcaatta ggagttaata acttcacggg aatagctaag ccgtcaaatg cagcttgcgt
     1381 gaaccctaat ttggagattc tcgacattca cgagaatcgc attaatggcg attttcccgc
     1441 gtggttgact gatttgacct ctttagtggt tttggatatt tctggaaacg gtttctccgg
     1501 tggtgttacg gctaaagttg ggaacttgat ggcgcttcag gagctgagag tagctaacaa
     1561 ttcacttgtt ggagaaattc cgacgagtat ccgtaattgc aaatcgttaa gagtcgttga
     1621 tttcgaagga aacaagttct cgggtcagat tccgggtttt ctaagtcagt tacggagcct
     1681 gacaacgata tctcttggaa gaaacgggtt ctcgggtcgg attccatcgg atcttctcag
     1741 cttgtacgga ctcgagacat tgaatctcaa tgaaaatcat ctcaccggag caataccgtc
     1801 ggagataaca aagctagcta atcttaccat ccttaatctc agtttcaaca gattctccgg
     1861 cgaagttccg tccaatgtgg gtgatctgaa aagcttgagt gttctcaaca taagtggatg
     1921 cgggttaacg ggtcggatac cggttagcat tagcggatta atgaagcttc aggttttgga
     1981 tataagtaaa cagagaatct ctggtcagtt accagtcgaa ctatttggtt taccggatct
     2041 tcaagtggtt gctcttggaa acaacttgct cggcggcgtc gtaccggaag gtttcagcag
     2101 cttggttagt cttaagtact taaatctcag ttccaactta ttctccggac atatcccaaa
     2161 gaactatgga tttctcaagt cattacaagt gctttcgttg tctcacaatc gaatatccgg
     2221 tacaatcccg ccggagattg ggaattgttc cagtcttgag gttcttgagc ttggttctaa
     2281 tagtttaaag ggtcacattc cggtttacgt ttccaagttg tctcttttga agaagcttga
     2341 tttgagtcac aacagtttaa cagggagtat tccggatcag atttctaagg attcttctct
     2401 agaatctctc ctgcttaatt cgaattctct atcgggtcga ataccggaat ctttatcaag
     2461 attgacgaat ctaaccgcgt tggatctttc gtcgaacaga ttgaattcca ctataccatc
     2521 aagtctctct cggcttcgtt tcttgaacta tttcaactta tcaagaaaca gtttggaagg
     2581 ggagattcca gaagctctag ctgcgcgttt cactaacccg accgtctttg tcaagaaccc
     2641 gggactctgc ggtaaaccgc ttggtataga atgtccaaac gttcgaagga gaagaagaag
     2701 aaagctactg gaggtgctgt tccagggacc aagcggcaat cccggcggcg atctggcccc
     2761 ccagatgctg cgcgagctgc aggagaccaa tgccgccctg caggacgtgc gcgagctgct
     2821 gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc gtgatggagt gcgacgcctg
     2881 cggcatgcag ccctctagag gctccggctc catcatccca gttgaggagg agaacccgga
     2941 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     3001 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     3061 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     3121 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     3181 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     3241 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     3301 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     3361 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     3421 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     3481 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     3541 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     3601 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     3661 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     3721 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     3781 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     3841 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     3901 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     3961 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     4021 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     4081 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     4141 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     4201 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     4261 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     4321 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggttc
     4381 tagaggaagc acctccggcg ataacgatta caaggatgat gatgataagg ccagcggcac
     4441 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     4501 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     4561 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4621 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4681 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     4741 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     4801 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4861 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4921 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5881 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5941 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6001 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6061 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6961 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     7021 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     7081 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     7141 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     7201 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7261 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     7321 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     7381 ggtaacgcca gggttttccc agtcacgacg tt
//
