LOCUS       sequence-298770-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G74360 X038_pECIA2 should be used with prey vector 
            AT1G74360 X038_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  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 6344)
  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: AT1G74360 X038_pECIA2
FEATURES             Location/Qualifiers
     source          1..6344
                     /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             2597..2620
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3326..3367
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3377..3394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3417..3434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3467..3601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3491..3510)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3545..3564
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3745..3761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3745..3761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3758..3780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3769..3785
                     /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(3793..3799,3800..3817,3818..3823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3838..3859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3976..3993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4147..4735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4227..4246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4906..5697,5698..5766))
                     /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     5529..5548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5767..5871)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5939..5957
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5995..6017)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6117..6136
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6330..6344,1..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 tctctggaca gtgatagaga
      541 agtcttgtta agcctaaaat cgtaccttga atcacggaac ccacagaacc gaggactata
      601 cacggaatgg aaaatggaga accaagatgt tgtgtgtcaa tggcctggaa tcatatgtac
      661 acctcagaga agtagagtta cagggatcaa tctaactgat tccaccattt caggtcctct
      721 attcaagaac ttctccgctt taacggagct cacatatctc gatttatcaa ggaatacgat
      781 cgaaggggag attccagatg acttgagccg ttgtcacaac ttaaagcatc tgaatctttc
      841 tcataatatc cttgaaggag agctaagttt acccgggttg tcaaatcttg aggttcttga
      901 tctttcgttg aataggatta cgggtgatat tcagtctagc ttcccattgt tctgcaacag
      961 tttggttgtt gcaaatctgt cgaccaacaa ctttactgga agaatcgatg acatcttcaa
     1021 cgggtgtagg aatctcaagt atgttgactt cagttccaat agattcagtg gagaagtgtg
     1081 gactggtttt gggaggctgg tcgagttttc agttgctgat aatcatctct ccgggaatat
     1141 ctctgcttca atgttcaggg ggaactgtac tttgcaaatg ttggatttgt ctgggaacgc
     1201 ttttggtggg gaatttccag gccaagtttc gaattgccag aatctaaatg tactgaatct
     1261 ttggggaaac aagttcacag ggaacattcc agctgagatt ggttccatat cctctctcaa
     1321 aggtttgtac ttggggaata atacattttc tcgagacata ccggaaactc tcctgaactt
     1381 gaccaacttg gtgtttctgg atttgagcag aaacaaattt ggaggagaca ttcaagaaat
     1441 ctttgggaga ttcacgcaag taaagtatct agtcttgcat gcaaactcat acgtaggagg
     1501 catcaattct tccaacatcc tcaagttacc caatctttca aggctagatt tgggctacaa
     1561 caatttctcg ggacagttac ctactgaaat ttctcagata cagagtttga agttcttgat
     1621 tcttgcttat aataacttca gtggcgatat accacaggag tatgggaaca tgccggggct
     1681 tcaagcactt gatctctcct ttaacaagct gaccggttcg ataccagctt catttgggaa
     1741 attgacctct cttttgtggc taatgcttgc aaacaactct ctatcaggag aaatccctcg
     1801 agagattggc aactgcacaa gccttttgtg gtttaacgtg gcaaacaacc agctctctgg
     1861 tagattccat cctgaattga ctagaatggg gagtaatcct tctccaacat ttgaagtgaa
     1921 taggcaaaac aaagacaaga taattgctgg ttctggtgaa tgcttggcga tgaagagatg
     1981 gattccagcg gagttccctc cgttcaactt tgtatacgca attctaacca aaaagagttg
     2041 cagaagccta tgggaccatg ttctcaaagg gtacggtctc tttcctgtct gttctgctgg
     2101 ttccacggta cgcacgctta aaatttcagc ctatcttcag ctcagtggaa ataagttttc
     2161 aggtgagatt cctgcaagca tctctcagat ggacaggctg agcacactac atttgggttt
     2221 caatgagttt gaggggaaac tgcctccaga gataggacaa ttgcctcttg cattcctcaa
     2281 ccttactcga aacaatttct cgggcgagat tcctcaagaa atcggaaatc tgaagtgcct
     2341 gcagaatctt gatctgtctt tcaacaattt ctctggaaac tttccaacga gtttgaatga
     2401 cttgaatgag ctgagtaagt tcaacatctc atataaccct ttcatttctg gcgcgatacc
     2461 aacaacagga caagtagcaa cctttgacaa agactccttt cttggaaatc cgctgctgcg
     2521 gtttcctagt ttcttcaacc aatcagggaa caacacgagg aagatttcca accaagtcct
     2581 aggaaacagg ccaagactgg aggtgctgtt ccagggacca gggagcgaca aaactcacac
     2641 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     2701 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     2761 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     2821 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     2881 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     2941 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     3001 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     3061 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     3121 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     3181 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     3241 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     3301 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3361 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3421 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3481 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3541 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3601 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3661 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     3721 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3781 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3841 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3901 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3961 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4021 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4081 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4561 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4981 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5041 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5101 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5161 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5221 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5281 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5341 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5401 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5461 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5521 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5581 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5641 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5701 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5761 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5881 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5941 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6001 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6061 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6121 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6181 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6241 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6301 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
