LOCUS       sequence-227206-        6662 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Prey vector AT1G05700 L9_pECIA14 should be used with bait vector 
            AT1G05700 L9_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6662)
  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 6662)
  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: AT1G05700 L9_pECIA14
FEATURES             Location/Qualifiers
     source          1..6662
                     /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             1958..1981
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3656..3679
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3695..3712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3735..3752)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3785..3919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3809..3828)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3863..3882
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4063..4079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4063..4079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4076..4098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4087..4103
                     /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(4111..4117,4118..4135,4136..4141))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4156..4177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4294..4311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4465..5053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4545..4564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5224..6015,6016..6084))
                     /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     5847..5866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6085..6189)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6257..6275
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6313..6335)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6435..6454
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6648..6662,1..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 tcaggtttca taagcatcga
      541 ttgtggaatt ccaagtggat catcgtacaa agacgacaca acaggtataa actacgtctc
      601 agattcatcc tttgtcgaaa ccggagtctc caaatcgatc cctttcacag ctcaaagaca
      661 gcttcaaaac ctgaggagct ttcccgaagg ttcaagaaac tgctatacat tgattccaat
      721 acaagggaag ggcaagaaat atctcataag agctagtttc atgtacggta actacgatgg
      781 agaaaatggt tcaccggagt ttgatctgtt tcttggtggc aatatttggg ataccgtctt
      841 gctcagcaat gggtcgagca ttgtttccaa agaagtcgtt tacttgagcc aatctgagaa
      901 catatttgtg tgtttgggaa acaaaggcaa aggaactcca tttatctcga ctttagagct
      961 taggtttctc gggaacgaca atacgacata tgattctcca aacggtgctc tcttcttctc
     1021 tagacgctgg gacttacgct ctctcatggg ttcacctgtt agatatgatg atgatgtgta
     1081 cgacagaatc tggatacctc gcaactttgg ctattgtaga gagatcaata cctcactccc
     1141 cgtgacctca gacaataata gttatagtct ttcaagtttg gtgatgagca cagcgatgac
     1201 tccaataaac acaacaagac ccatcacaat gactttggaa aacagtgatc caaatgtcag
     1261 gtactttgtc tacatgcact tcgcagaggt tgaagatctt agtctcaaac cgaaccagac
     1321 aagagagttc gacatcagca ttaacggagt gacagttgct gctggattca gccccaagta
     1381 tcttcagaca aacacgtttt ttctaaaccc tgagagccaa tcgaaaattg cgttttcact
     1441 cgttcgaacc ccaaagtcca ctcttccgcc aattgttaac gctctagaga tctacgtcgc
     1501 aaatagtttc tcgcagtctc tcactaacca agaggatggt gacgcggtta cgagtctaaa
     1561 gacgagttat aaagtgaaga agaactggca cggagatcct tgtttgccta atgactacat
     1621 ttgggagggc cttaattgca gttatgatag tcttactcct ccaagaatca catcactgaa
     1681 cttatcatca agcgggttaa cgggtcatat atcttcctcc ttctctaacc tcacaatgat
     1741 tcaagagcta gacttatcaa acaatggctt aaccggagac attccggagt tcctctcaaa
     1801 actgaagttc ttgagggttt taaatctaga aaataacact cttaccggtt cagttccatc
     1861 cgagttatta gaaagatcaa acaccggatc attctcgcta aggctaggag aaaatccagg
     1921 actctgtact gagatttctt gcagaaaaag caacagcctg gaggtgctgt tccagggacc
     1981 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     2041 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     2101 gaagaacacc gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc
     2161 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     2221 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     2281 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     2341 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     2401 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     2461 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     2521 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     2581 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     2641 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     2701 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     2761 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     2821 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     2881 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     2941 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     3001 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     3061 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     3121 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     3181 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     3241 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     3301 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     3361 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     3421 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     3481 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     3541 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     3601 cggcaccacc gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta
     3661 caaggatgat gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta
     3721 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     3781 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3841 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3901 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3961 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     4021 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4081 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4141 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4201 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4381 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5281 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5341 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5401 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5461 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5521 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5581 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5641 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5701 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5761 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5821 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5881 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5941 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6001 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6061 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6181 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6241 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6301 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6361 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6421 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6541 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6601 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6661 tt
//
