LOCUS       sequence-274570-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G24650 X034_pECIA14 should be used with bait vector 
            AT1G24650 X034_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  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 6533)
  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: AT1G24650 X034_pECIA14
FEATURES             Location/Qualifiers
     source          1..6533
                     /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             1829..1852
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3527..3550
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3566..3583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3606..3623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3656..3790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3680..3699)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3734..3753
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3934..3950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3934..3950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3947..3969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3958..3974
                     /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(3982..3988,3989..4006,4007..4012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4027..4048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4165..4182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4336..4924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4416..4435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5095..5886,5887..5955))
                     /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     5718..5737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5956..6060)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6128..6146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6184..6206)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6306..6325
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6519..6533,1..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 agccctgatg aggcggtcat
      541 gatagctctg agagactcct tgaagctatc tggaaatcca aactggtccg ggtcggatcc
      601 ttgtaagtgg tctatgttca taaaatgcga cgcaagcaat cgggtcaccg cgatccaaat
      661 cggggacaga ggaatctcag gaaagctacc tcctgatctc ggaaagctaa catctttgac
      721 gaagttcgag gtcatgagga accgtttaac aggaccaata ccgagtcttg ctgggttgaa
      781 atcgttggta actgtgtatg ctaatgacaa tgatttcact tctgtccctg aggatttctt
      841 ctctggtttg agttctcttc agcatgtttc tctggataac aatccttttg attcttgggt
      901 gatacctccg agtttggaaa acgcgacttc ccttgtagat ttctctgctg tcaactgcaa
      961 tctttccggg aagatcccgg attatctctt cgaggggaaa gatttctcta gtttgacgac
     1021 tctgaaactg tcttataact cgcttgtatg tgaatttccc atgaatttct ccgactcacg
     1081 tgttcaagtt ctgatgctta acgggcaaaa gggaagggag aagcttcatg ggtcaatttc
     1141 gtttctgcaa aagatgactt ctttgaccaa tgttacgctg caagggaaca gcttctccgg
     1201 tcctttacca gacttttctg gtttagtctc gttgaaaagt ttcaatgtta gggagaatca
     1261 gctttcgggt ctcgttccct cttctttgtt tgagctgcaa tctctttctg atgttgcttt
     1321 aggtaataac ttgcttcaag gaccaactcc aaacttcacg gctcctgaca ttaagcctga
     1381 tctgaatggt ctcaatagtt tctgcttaga tactccaggt acttcttgtg atccccgtgt
     1441 caacacgttg ctttcaatcg ttgaagcctt tggttacccg gtgaattttg cggagaaatg
     1501 gaaaggaaat gatccttgta gtggatgggt agggatcact tgtactggaa ctgatatcac
     1561 agttatcaac ttcaaaaact tggggctcaa cggcactata tctccacgat ttgctgattt
     1621 tgcgtctctt cgtgttatta atctctctca gaataatctt aatggtacta ttccacaaga
     1681 gctcgctaag ttgagtaact tgaaaacact ggatgtttcg aagaatcggt tgtgtggtga
     1741 agtacctcgt tttaacacaa ctattgttaa cacgactggg aattttgagg actgtcctaa
     1801 tggcaatgct ggaaagaaag catctagtct ggaggtgctg ttccagggac caagcggcaa
     1861 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     1921 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     1981 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2041 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2101 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     2161 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     2221 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2281 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2341 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2401 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     2461 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     2521 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     2581 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     2641 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     2701 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     2761 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     2821 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     2881 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     2941 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3001 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3061 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3121 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     3181 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3241 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3301 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3361 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3421 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     3481 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     3541 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3601 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3661 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3721 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3781 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3841 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     3901 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3961 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4021 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4081 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4141 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4621 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4981 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5041 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5101 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5161 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5221 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5281 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5341 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5401 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5461 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5521 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5581 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5641 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5701 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5761 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5821 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5881 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5941 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6061 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6121 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6181 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6241 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6301 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6361 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6421 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6481 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
