LOCUS       sequence-274561-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G37450 X025_pECIA14 should be used with bait vector 
            AT5G37450 X025_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  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 6812)
  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: AT5G37450 X025_pECIA14
FEATURES             Location/Qualifiers
     source          1..6812
                     /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             2108..2131
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3806..3829
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3845..3862
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3885..3902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3935..4069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3959..3978)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4013..4032
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4213..4229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4213..4229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4226..4248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4237..4253
                     /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(4261..4267,4268..4285,4286..4291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4306..4327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4444..4461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4615..5203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4695..4714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5374..6165,6166..6234))
                     /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     5997..6016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6235..6339)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6407..6425
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6463..6485)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6585..6604
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6798..6812,1..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 actcatccca ctgatgttag
      541 tgctctgcag tatgttcacc gcaaactaaa ggaccctctg aatcatcttc aagactggaa
      601 aaagacggat ccatgcgcct ctaactggac cggtgtcatc tgcatccctg atccttctga
      661 tggctttctt catgttaaag aattgcggtt gctaaacatg aacctcactg gacaactggc
      721 acctgaactc ggcttactat caaatcttac catattgaac ttcatgtgga acgatctcac
      781 cggtcagatt ccaccagaac taggaaactt aactcatctc atcttcttgc tattaagtgg
      841 aaatcaactg acaggatctt tgcctcaaga actcggttct ctttccaatc tcttaatcct
      901 tcagatagat tacaatgaaa ttagcggaaa acttccaacg tccttggcaa atttgaaaaa
      961 actgaagcat tttcacatga acaataactc aatcacgggt cagattccgc ccgagtactc
     1021 taccttaacc aatgttctgc acttcttgat ggacaataac aagctaactg gtaatcttcc
     1081 accggagctc gctcagatgc caagcttgag aattctacaa ttggacggta gcaactttga
     1141 tggcacagag atcccatctt cttacggatc gataccaaat ctcgtcaaat tgagtctcag
     1201 aaactgcaac ctagaaggac caattcctga tttgagcaaa tcactggttc tctattactt
     1261 agatatttcc tccaacaaac tcacaggaga aattcccaag aacaagttct ctgcaaacat
     1321 cacaactata aacttgtaca acaatttgct cagcggatcc attccttcta atttttcagg
     1381 ccttccacgg ttacagagac tgcaagtgca gaacaacaac ttgagtgggg agattcctgt
     1441 gatatgggaa aataggatct tgaaggcaga ggaaaaactt atcttggact tgaggaacaa
     1501 catgttttca aatgtgtcaa gtgttcttct aaaccctccc tccaatgtca cagtcaagct
     1561 ttacggaaac ccggtttgtg caaatgttaa tgcgggaaag ttagctgatc tctgtggtat
     1621 ctcaacgtta gaagtggaat ctcccgcaac ttcttcggaa accatctcga cgggagactg
     1681 caaacgtcag tcgtgccccg taagcgaaaa ctatgactac gtgattgggt ctccagttgc
     1741 atgtttctgc gctgcaccac ttggcattga tcttcggctt aggagtccaa gtttctcgga
     1801 ctttcgacct tacaaagtct cctacatgct cgatgtagca tccccgaaaa atctcgggat
     1861 aaacccgtat cagatatcta ttgatacatt tgcatggcag tcgggtccaa ggctattcat
     1921 gaacatgaag attttccctg agtacagtga attaaacagt aaattcaaca gcacagaggt
     1981 tcaacgtatt gtggacttct ttgcaacttt cactctgaat actgatgatt ctcttggccc
     2041 ttatgagatt attagtatca acactggtgc ttacaaagac gttactatca tattcccgaa
     2101 aaagtcgctg gaggtgctgt tccagggacc aagcggcaat cccggcggcg atctggcccc
     2161 ccagatgctg cgcgagctgc aggagaccaa tgccgccctg caggacgtgc gcgagctgct
     2221 gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc gtgatggagt gcgacgcctg
     2281 cggcatgcag ccctctagag gctccggctc catcatccca gttgaggagg agaacccgga
     2341 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     2401 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     2461 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     2521 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     2581 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     2641 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     2701 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     2761 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     2821 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     2881 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     2941 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     3001 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     3061 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     3121 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     3181 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     3241 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     3301 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     3361 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     3421 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     3481 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     3541 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     3601 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     3661 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     3721 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggttc
     3781 tagaggaagc acctccggcg ataacgatta caaggatgat gatgataagg ccagcggcac
     3841 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3901 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3961 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4021 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4081 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     4141 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     4201 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4261 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4321 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6361 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6421 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6481 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6541 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6601 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6661 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6721 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6781 ggtaacgcca gggttttccc agtcacgacg tt
//
