LOCUS       sequence-270303-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G43020 K6_pECIA14 should be used with bait vector 
            AT5G43020 K6_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  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 5969)
  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: AT5G43020 K6_pECIA14
FEATURES             Location/Qualifiers
     source          1..5969
                     /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             1265..1288
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2963..2986
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3002..3019
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3042..3059)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3092..3226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3170..3189
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3370..3386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3370..3386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3383..3405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3394..3410
                     /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(3418..3424,3425..3442,3443..3448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3463..3484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3601..3618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3772..4360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3852..3871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4531..5322,5323..5391))
                     /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     5154..5173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5392..5496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5564..5582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5620..5642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5742..5761
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5955..5969,1..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 tccaaccaca caaagcgcgt
      541 gtttcattct caccgtgacg tttctgcttt gttacgtttt aaatccaaag ctgatttatg
      601 gaacaagata aacacgagct ctcatttttg tcaatggtgg ggtgttacgt gttatggaaa
      661 cagagttgtt cgtttggtta ttgaagatct ttatcttgga gggcgactca ttcctgactc
      721 agttaataaa ctcgaccagc ttcgagttct gagtctcaag aacacttcct taaccggacc
      781 gttaccagat ttctccggtt tggttaatct caaatcatta ttcctcgatc acaactcttt
      841 ttccggttcg tttcctctct cggttctcgc ttttcaccgg ttaagaactc tcgatttctc
      901 cttcaataac ctcaccggtc caatcccatc cggtttagtt ctctccgacc ggttgattta
      961 tctccgactt gattcgaatc ggttcaacgg tcccgtcccg cctttaaacc aatccacgct
     1021 tcatacattc aatgtttcgg ttaacaactt aaccggtgct gtaccggtga caaccgttct
     1081 gttacgattc ggtatttctt catttctcaa aaaccctaat ctctgcggtg agattgttca
     1141 caaagaatgc aatccgagag ccaaattctt cactccggta acggcggcgc cgtcaccgaa
     1201 gatggttctc ggtcaaatag cgcaaatagg cggcgcaaga ttatcccggc caagtcaaaa
     1261 caaactggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     1321 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     1381 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     1441 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     1501 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     1561 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     1621 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     1681 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     1741 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     1801 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     1861 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     1921 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     1981 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     2041 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     2101 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     2161 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     2221 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     2281 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     2341 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     2401 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     2461 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     2521 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     2581 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     2641 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     2701 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     2761 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     2821 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     2881 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     2941 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     3001 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     3061 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     3121 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3181 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3241 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3301 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     3361 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4201 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4321 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4501 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4561 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4621 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4681 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4741 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4801 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4861 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4921 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4981 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5041 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5101 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5161 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5221 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5281 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5341 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5401 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5461 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5521 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5581 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5641 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5701 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5761 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5821 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5881 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5941 aacgccaggg ttttcccagt cacgacgtt
//
