LOCUS       sequence-352626-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G63430 O10_pECIA14 should be used with bait vector 
            AT1G63430 O10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  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 6014)
  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: AT1G63430 O10_pECIA14
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..427
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     445..498
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1288..1311
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2986..3009
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3025..3042
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3065..3082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3115..3249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3139..3158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3193..3212
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3393..3409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3393..3409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3406..3428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3417..3433
                     /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(3441..3447,3448..3465,3466..3471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3486..3507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3624..3641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3795..4383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3875..3894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4554..5345,5346..5414))
                     /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     5177..5196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5415..5519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5587..5605
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5643..5665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5765..5784
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5978..6000
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5993..6009
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgttgc aggacaggat gtggtgcccg atgtgactag ctctttgctg caggccgtcc
       61 tatcctctgg ttccgataag agacccagaa ctccggcccc ccaccgccca ccgccacccc
      121 catacatatg tggtacgcaa gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag
      181 ctttgcatcc catacaagtc cccaaagtgg agaaccgaac caattcttcg cgggcagaac
      241 aaaagcttct gcacacgtct ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt
      301 gaatcgaacg aaagacccgt gtgtaaagcc gcgtttccaa aatgtataaa accgagagca
      361 tctggccaat gtgcatcagt tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa
      421 ctaaaggggg gatccgatct caatatgaag ttatgcatat tactggccgt cgtggccttt
      481 gttggcctct cgctcggggg gtttgcctct aatgaagttc aagcacttag aagattcaag
      541 gaagctatct atgaggaccc tttgctagtt atgtctaatt ggaatgaccc caattcagat
      601 ccttgtgatt ggaccggcat ttattgctct ccatctaaag atcatgttat caagataaat
      661 atatccgctt catcgataaa agggtttctt gcgccggaat tgggtcaaat aacgtacctg
      721 caggaactaa tcctacatgg gaacattcta atagggacaa tacctaagga gataggaaac
      781 ttaaagaatc tcaaaatctt ggacttggga aacaatcatc ttatgggacc gatccctgca
      841 gagatcggga gtttgtccgg catcatgata atcaaccttc agtccaatgg tttgactgga
      901 aagttgcctg cagagctcgg taacttgaag taccttagag aacttcatat tgacaggaat
      961 aggcttcaag gaagtcttct tgttgctgga gcatcgggct atcagtcgaa agtgtattct
     1021 tcaaattcaa gtgcaaacat cgctggtttg tgcaaatctt taaaagtagc tgacttttca
     1081 tacaactttt tcgtgggaaa tattccaaaa tgtttagaaa accttccaag gacgagcttt
     1141 caagggaact gcatgcaaaa caaggatctt aagcatagat cttcttccca atgcgctaat
     1201 gcacaactgg tcaaaactca tggaagcccg agcgctgctc ctaagcacca atcagctcaa
     1261 atggtggcta agcaccatag agcatcactg gaggtgctgt tccagggacc aagcggcaat
     1321 cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa tgccgccctg
     1381 caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc
     1441 gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc catcatccca
     1501 gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct gggtgccgcc
     1561 aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct gggcgatggg
     1621 atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa ggacaaactg
     1681 gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc caagacatac
     1741 aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct gtgcggggtc
     1801 aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca gtgcaacacg
     1861 acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg
     1921 ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta cgcccacacg
     1981 gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc
     2041 caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct aggtggaggc
     2101 cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga ctacagccaa
     2161 ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt
     2221 gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc gtctgtgacc
     2281 catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg agactccaca
     2341 ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag caggaacccc
     2401 cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca tgaaagcagg
     2461 gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag ggcgggccag
     2521 ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca cgtcttctcc
     2581 ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg caaggcccgg
     2641 gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt gctcaaggac
     2701 ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg gcagcagtca
     2761 gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc
     2821 ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca cgtcatggcc
     2881 ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc
     2941 gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta caaggatgat
     3001 gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta aacccgctga
     3061 tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca
     3121 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3181 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3241 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     3301 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg
     3361 acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3421 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3481 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3541 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3601 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3661 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3721 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3781 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3841 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3901 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3961 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4021 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4081 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4141 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4201 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4261 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4321 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4381 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4441 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4501 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4561 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4621 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4681 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4741 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4801 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4861 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4921 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4981 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5041 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5101 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5161 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5221 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5281 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5341 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5401 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5461 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5521 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5581 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5641 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5701 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5761 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5821 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5881 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5941 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     6001 acggccagtg aatt
//
