LOCUS       sequence-297036-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G53420 X068_pECIA2 should be used with prey vector 
            AT1G53420 X068_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  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 5954)
  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: AT1G53420 X068_pECIA2
FEATURES             Location/Qualifiers
     source          1..5954
                     /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             2207..2230
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2936..2977
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2987..3004
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3027..3044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3077..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3155..3174
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3355..3371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3355..3371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3368..3390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3379..3395
                     /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(3403..3409,3410..3427,3428..3433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3448..3469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3586..3603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3837..3856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4516..5307,5308..5376))
                     /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     5139..5158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5377..5481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5549..5567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5605..5627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5727..5746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5940..5954,1..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 tctgcaactt tgccaacaca
      541 agaaggggag gcttttaaag ttgtactaac cacattgaaa aagacgaata ttgatttaaa
      601 tgtggatcca tgcgaagtct catctactgg aaatgaatgg tcaaccatca gcaggtggaa
      661 agacctagta acttgtgact gctccttcgt taatggaacc atttgccaca ttaccaaaat
      721 aaatttgaag agagaaaatc ttcaagggtc tcttccaaaa gagttagtgg gacttccttt
      781 actgcaagag attgatctct caagaaacta tctcaatggt tccatccctc ctgaatgggg
      841 agtcttgcca cttgtcaaca tttggcttct cggaaaccga ttaacaggtc caatccctaa
      901 ggagtttgga aacattacaa ctcttactag ccttgtcttg gaagccaatc aactttcagg
      961 agaattacct cttgagctag ggaatctacc aaacattcaa caaatgattc ttagctcgaa
     1021 taactttaat ggtgaaattc catctacatt tgcaaaacta accaccttga gagattttcg
     1081 tgtaagtgac aaccagttgt caggaacaat tccagatttc atccaaaaat ggactaagct
     1141 tgagcgactg tttattcaag caagtggttt agttggacca attcccattg ccattgcttc
     1201 tctcgtagag ttaaaagact tgagaatcag cgatttgaat ggacccgaat ctccatttcc
     1261 acagttaagg aacataaaaa agatggagac attaattctt aggaactgca atctcacagg
     1321 agacttacct gattatcttg gaaagattac ctccttcaaa ttcttagatc ttagcttcaa
     1381 taaattaagt ggagccatcc ctaacactta tattaatctt agagacggag gttacatata
     1441 ttttacagga aacatgttaa acgggtcggt tccagattgg atggtaaata aaggatataa
     1501 aattgatctt agttacaaca atttctcagt agatccaacc aatgcagtat gcaagtacaa
     1561 taatgtactt tcatgcatga gaaactacca atgtcctaaa actttcaatg ctcttcatat
     1621 aaactgtggt ggggatgaaa tgtctatcaa tgggacaata tatgaatctg ataagtacga
     1681 tagactggag agttggtacg aaagtcgaaa tggatggttc tcaaacaacg taggagtctt
     1741 cgtagacgat aaacatgttc ccgagagagt aaccatcgag tctaactcat ccgagctcaa
     1801 tgtggttgat tttggtctgt atacgcaagc tcgcatatca gctatctcac tcacttacta
     1861 tgcattgtgt ttagaaaatg gaaattacaa cgttaatctc catttcgccg agattatgtt
     1921 caatggtaac aacaattatc aaagcttggg tagacggttt ttcgacatat acattcagag
     1981 gaagctcgag gttaaagatt tcaatatcgc caaggaggca aaagatgttg gaaacgttgt
     2041 tattaagaca tttccggtcg agataaaaga tggaaagttg gagatacgat tgtattgggc
     2101 cgggagagga actacagtca ttcccaaaga acgtgtttat ggtcctctca tatcagctat
     2161 atcagtggat tcaagtgtta atccatctcc tagaaatggt atgagtctgg aggtgctgtt
     2221 ccagggacca gggagcgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct
     2281 ggggggaccg tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg
     2341 gacccctgag gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt
     2401 caactggtac gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca
     2461 gtacaacagc acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa
     2521 tggcaaggag tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac
     2581 catctccaaa gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg
     2641 ggaggagatg accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag
     2701 cgacatcgcc gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc
     2761 tcccgtgctg gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag
     2821 caggtggcag caggggaacg tcttctcatg ctccgtgatg catgagggtc tgcacaacca
     2881 ctacacgcag aagagcctct ccctgtctcc gggtaaatct agagggccct tcgaaggtaa
     2941 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     3001 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     3061 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3121 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3181 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3241 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3301 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     3361 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3421 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5521 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5581 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5641 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5701 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5761 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5821 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5881 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5941 ccagtcacga cgtt
//
