LOCUS       sequence-288222-        7076 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Prey vector AT3G47570 L7_pECIA14 should be used with bait vector 
            AT3G47570 L7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7076)
  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 7076)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AT3G47570 L7_pECIA14
FEATURES             Location/Qualifiers
     source          1..7076
                     /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             2372..2395
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4070..4093
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4109..4126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4149..4166)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4199..4333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4223..4242)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4277..4296
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4477..4493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4477..4493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4490..4512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4501..4517
                     /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(4525..4531,4532..4549,4550..4555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4570..4591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4708..4725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4879..5467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4959..4978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5638..6429,6430..6498))
                     /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     6261..6280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6499..6603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6671..6689
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6727..6749)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6849..6868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7062..7076,1..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 accgatgaaa ctgataggca
      541 agctttgctt cagttcaagt ctcaagtttc tgaagacaaa agagttgtct tgtcctcatg
      601 gaaccactca ttcccactct gcaattggaa gggcgttaca tgtggccgta aaaacaaaag
      661 agttactcat ctggaactcg gaagattgca actaggcgga gtgatatcac catccattgg
      721 taatctatcg tttcttgtat cacttgatct ctatgaaaac ttttttggtg gtactattcc
      781 tcaagaggtg ggacagttgt ctagactcga atacttggat atgggcataa attatcttag
      841 gggaccgatt ccattaggtc tatacaattg ctctagattg ttgaaccttc gtttagattc
      901 aaatcgtctt gggggaagtg ttccttcaga gctaggatca ttgacgaacc ttgttcagtt
      961 aaatctttat ggaaacaata tgagaggaaa gctccctaca tcactaggaa acttgacgtt
     1021 actagaacaa cttgcattgt cgcataacaa tctggaagga gaaattccga gtgatgtagc
     1081 tcaactgact caaatatgga gtcttcaatt agtagccaac aatttctccg gtgtttttcc
     1141 tcccgctctc tacaacttgt cctcacttaa gcttctaggt atcggttata atcatttctc
     1201 gggtcgcctt agacctgact taggtattct actaccaaat cttctatcgt ttaatatggg
     1261 agggaattat ttcacagggt ccattccaac aactctttcc aatatctcta ctcttgaaag
     1321 attgggtatg aatgagaaca atctgacagg aagtattcct acattcggaa atgttccaaa
     1381 tttaaaattg ttatttcttc ataccaattc tttaggaagt gattcttcta gagatcttga
     1441 attcctcact tctttgacaa actgcaccca actagagacc ttagggattg gcagaaatag
     1501 gcttgggggt gacttgccta tatctattgc gaacctatcc gcaaaactcg taactctaga
     1561 ccttggagga accctcattt ccggaagcat tccttatgac attggaaatc tcataaacct
     1621 acaaaagctt attttagatc aaaatatgtt gagtggacca ctcccaacct ctcttggaaa
     1681 gcttttgaat ttgaggtatt taagtctatt ttcaaataga ttatcaggag gtataccagc
     1741 ttttataggc aacatgacta tgttagaaac acttgatttg tccaacaatg gttttgaagg
     1801 tattgttcct acaagtcttg gcaactgtag tcacctacta gaattgtgga ttggggataa
     1861 taagttgaat gggactatac ctctggaaat tatgaaaatt cagcagcttc ttcgcctaga
     1921 catgtcagga aattctttga tcggctctct cccgcaggat atcggagcgc tacaaaatct
     1981 tggtacacta tcacttgggg ataataaatt atcagggaaa cttccacaaa ctttgggaaa
     2041 ctgtcttaca atggaaagtc tttttttgga aggaaatttg ttttatggag atattccaga
     2101 tttaaaaggg ttggtgggtg ttaaagaagt tgatttgtct aacaatgatc tctctggaag
     2161 tatacctgaa tactttgcaa gcttttccaa gttggagtat ctcaatctat cttttaacaa
     2221 tcttgaggga aaggtgccag tgaaaggaat atttgagaat gctactacag tttccatagt
     2281 tgggaacaat gatttatgtg gaggtattat ggggtttcaa ctaaagccat gcctttcgca
     2341 agcaccatct gtggtgaaga agcattcatc tctggaggtg ctgttccagg gaccaagcgg
     2401 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     2461 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     2521 caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg gctccatcat
     2581 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     2641 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     2701 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     2761 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2821 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2881 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2941 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     3001 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     3061 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     3121 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     3181 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     3241 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     3301 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     3361 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     3421 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     3481 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     3541 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     3601 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     3661 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     3721 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3781 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3841 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3901 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3961 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     4021 caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg attacaagga
     4081 tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag tttaaacccg
     4141 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     4201 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4261 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4321 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4381 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     4441 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4501 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4561 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4621 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6601 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6661 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6721 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6781 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6841 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6901 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6961 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7021 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
