LOCUS       sequence-270219-        5819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G10290 I2_pECIA14 should be used with bait vector 
            AT5G10290 I2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5819)
  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 5819)
  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: AT5G10290 I2_pECIA14
FEATURES             Location/Qualifiers
     source          1..5819
                     /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             1115..1138
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2813..2836
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2852..2869
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2892..2909)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2942..3076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2966..2985)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3020..3039
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3220..3236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3220..3236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3233..3255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3244..3260
                     /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(3268..3274,3275..3292,3293..3298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3313..3334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3451..3468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3622..4210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3702..3721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4381..5172,5173..5241))
                     /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     5004..5023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5242..5346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5414..5432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5470..5492)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5592..5611
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5805..5819,1..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 tctccagatg ctcaagggga
      541 tgcactgttt gcgttgagga tctccttacg tgcattaccg aatcagctaa gtgactggaa
      601 tcagaaccaa gttaatcctt gcacttggtc ccaagttatt tgtgatgaca aaaactttgt
      661 cacttctctt acattgtcag atatgaactt ctcgggaacc ttgtcttcaa gagtaggaat
      721 cctagaaaat ctcaagactc ttactttaaa gggaaatgga attacgggtg aaataccaga
      781 agactttgga aatctgacta gcttgactag tttggatttg gaggacaatc agctaactgg
      841 tcgtatacca tccactatcg gtaatctcaa gaaacttcag ttcttgacct tgagtaggaa
      901 caaacttaat gggactattc cggagtcact cactggtctt ccaaacctgt taaacctgct
      961 gcttgattcc aatagtctca gtggtcagat tcctcaaagt ctgtttgaga tcccaaaata
     1021 taatttcacg tcaaacaact tgaattgtgg cggtcgtcaa cctcaccctt gtgtatccgc
     1081 ggttgcccat tcaggtgatt caagcaagcc taaactggag gtgctgttcc agggaccaag
     1141 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc
     1201 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa
     1261 gaacaccgtg atggagtgcg acgcctgcgg catgcagccc tctagaggct ccggctccat
     1321 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     1381 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     1441 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     1501 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     1561 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     1621 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     1681 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     1741 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     1801 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     1861 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     1921 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     1981 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     2041 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     2101 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     2161 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     2221 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     2281 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     2341 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     2401 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     2461 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     2521 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     2581 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     2641 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     2701 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     2761 caccaccgac gccgcgcacc cgggttctag aggaagcacc tccggcgata acgattacaa
     2821 ggatgatgat gataaggcca gcggcaccgg tcatcatcac catcaccatt gagtttaaac
     2881 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     2941 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3001 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3061 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3121 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     3181 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3241 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3301 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4021 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4141 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4261 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4321 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4381 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4441 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4501 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4561 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4621 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4681 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4741 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4801 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4861 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4921 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4981 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5041 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5101 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5161 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5221 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5281 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5341 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5401 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5461 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5521 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5581 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5641 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5701 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5761 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
