LOCUS       sequence-270073-        6638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G07550 L10_pECIA14 should be used with bait vector 
            AT1G07550 L10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6638)
  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 6638)
  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: AT1G07550 L10_pECIA14
FEATURES             Location/Qualifiers
     source          1..6638
                     /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             1934..1957
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3632..3655
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3671..3688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3711..3728)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3761..3895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3785..3804)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3839..3858
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4039..4055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4039..4055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4052..4074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4063..4079
                     /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(4087..4093,4094..4111,4112..4117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4132..4153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4270..4287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4441..5029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4521..4540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6233..6251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6289..6311)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6411..6430
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6624..6638,1..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 agtttggatt gtggattagc
      541 ttctaacgag tctccttaca acgaagcaaa tagcaatctg acgtacatat ccgatgctga
      601 tttcattcaa ggaggaaaaa ctggtaatgt ccaaaaagac ttgcttatga aactcaggaa
      661 gccatatacg gttttgagat actttccgga tggaatccga aattgttaca gtctaaatgt
      721 caagcaagac acaaattatc taatcagggt tatgtttcga tatggaaact atgacgggct
      781 taacaattct ccaagattcg acctctatct tggtcctaat atttggacaa ccatagatat
      841 ggggaagtca ggtgatggag ttctggagga gatcattcac atcacaaggt ctaacatctt
      901 ggacatatgt ctcgtcaaga cagggacgag tacaccaatg atatcatcca tagagttgag
      961 acccttgcta tatgatactt atattgctca aaccggttcc ttgaggaatt ataaccggtt
     1021 ctatttcacc gattcaaaca actatatacg ttatccccaa gatgtgcatg atagaatttg
     1081 ggtgccgttg atattaccag aatggacgca tataaacaca agtcaccacg ttattgattc
     1141 tatagatggc tatgacccac cacaagatgt acttaggacc ggcgccatgc ccgctaatgc
     1201 cagtgatcca atgaccatta cctggaattt gaagacggct actgaccagg tttacggtta
     1261 tatttacatc gccgagatca tggaagtcca ggccaatgaa actagagaat tcgaagttgt
     1321 ggtaaataat aaagttcatt tcgatccttt ccgtcctacc agatttgaag cacaagttat
     1381 gttcaacaat gtaccactga catgcgaggg aggattttgt cgtctgcaac ttataaaaac
     1441 tccgaaatcg actcttccac ccttgatgaa tgctttcgaa attttcacgg gcattgagtt
     1501 tcctcaatca gaaacaaatc aaaatgatgt cattgctgta aagaatattc aagctagtta
     1561 tggactgaac agaatcagct ggcagggcga tccatgtgtc cctaaacagt ttttgtggac
     1621 tggcttaagc tgcaacgtaa ttgatgtgtc cacaccacca agaatcgtga aattagattt
     1681 gtcttcaagt ggattaaatg gagttatacc acctagcata caaaatctaa ctcaacttca
     1741 agaactggac ctgtcccaaa acaacttgac tggtaaagtg cctgaatttc tagccaagat
     1801 gaaatatttg ttggtcataa acttaagtgg aaacaaactt agcggtttgg ttcctcaagc
     1861 ccttcttgat agaaaaaagg aaggattaaa gctattggtt gacgaaaata tgatttgtgt
     1921 gtcttgcggt acactggagg tgctgttcca gggaccaagc ggcaatcccg gcggcgatct
     1981 ggccccccag atgctgcgcg agctgcagga gaccaatgcc gccctgcagg acgtgcgcga
     2041 gctgctgcgc caccgcgtga aggagatcac cttcctgaag aacaccgtga tggagtgcga
     2101 cgcctgcggc atgcagccct ctagaggctc cggctccatc atcccagttg aggaggagaa
     2161 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc
     2221 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac
     2281 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc
     2341 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca
     2401 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca
     2461 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga
     2521 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac
     2581 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg
     2641 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac
     2701 gcagctcatc tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt
     2761 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct
     2821 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg
     2881 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct
     2941 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct
     3001 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct
     3061 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact
     3121 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga
     3181 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc
     3241 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta
     3301 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga
     3361 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga
     3421 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct
     3481 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct
     3541 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc
     3601 gggttctaga ggaagcacct ccggcgataa cgattacaag gatgatgatg ataaggccag
     3661 cggcaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     3721 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     3781 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3841 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3901 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3961 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     4021 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4081 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4141 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6241 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6301 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6361 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6421 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6481 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6541 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6601 aagttgggta acgccagggt tttcccagtc acgacgtt
//
