LOCUS       sequence-274548-        6983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G14840 X033_pECIA14 should be used with bait vector 
            AT3G14840 X033_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6983)
  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 6983)
  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: AT3G14840 X033_pECIA14
FEATURES             Location/Qualifiers
     source          1..6983
                     /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             2279..2302
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3977..4000
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4016..4033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4056..4073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4106..4240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4130..4149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4184..4203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4384..4400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4384..4400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4397..4419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4408..4424
                     /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(4432..4438,4439..4456,4457..4462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4477..4498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4615..4632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4786..5374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4866..4885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5545..6336,6337..6405))
                     /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     6168..6187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6406..6510)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6578..6596
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6634..6656)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6756..6775
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6969..6983,1..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 tctgcaacat taccaaaaga
      541 agaagtggat gctttacaaa gcgtcgcgac ggcgttgaag aagagtaatt ggaatttcag
      601 tgtggatcca tgtgacgaaa ccttgtctga aggcggatgg aggaatccta acgccgctaa
      661 gggattcgaa gacgccgtta cttgtaactg ctcctccgtc atctgccacg tcaccaacat
      721 agttctcaag gcacaagacc ttcaaggatc tcttcctaca gatctttcag gacttccttt
      781 cttgcaagag cttgatctga ctagaaacta tctcaacggt tccattcctc ccgaatgggg
      841 agcctcatca cttctaaaca tctcactact tggaaaccgg ataagtggtt caatcccaaa
      901 agaacttgga aaccttacaa ccctttctgg tcttgtcttg gaatacaacc aactctcagg
      961 gaagatacct ccagagctcg ggaatctacc aaaccttaaa agactgttac ttagctcgaa
     1021 taacttgagc ggggaaatcc caagtacatt cgccaaactt actacgttga ctgatttacg
     1081 tattagtgac aaccagttca ctggtgctat accagatttc atccagaact ggaaaggact
     1141 agagaaactg gttattcaag caagtggttt agttggaccg attcctagtg ccattggtct
     1201 tctgggaacc ttgacagact tgagaatcac tgacttgagt ggacctgaat ctccatttcc
     1261 gccactaaga aacatgacat cgatgaagta tttgattctt aggaactgta accttacagg
     1321 agacttacct gcgtatcttg gacagaatag aaaattaaag aatctagatc ttagctttaa
     1381 caaactgagt ggaccaatcc cagccacata ttccggtctt tcagatgtag atttcatata
     1441 ctttacaagt aacatgttaa atggacaagt accaagttgg atggtagacc aaggagacac
     1501 gattgatatt acttacaata acttctccaa agataaaacc gaagaatgtc aacaaaaatc
     1561 cgtgaacacg ttttcaagca caagcccttt agtggcaaat aactcctcaa atgtttcgtg
     1621 tctgagtaaa tatacctgtc ctaaaacttt ctacggcctt catataaact gtggtggtaa
     1681 tgaaataaca agcaatgaga ctaagtatga tgctgacaca tgggatacac cgggttacta
     1741 tgatagtaaa aacgggtggg tttctagcaa cacgggaaac ttcttggatg atgatcggac
     1801 taacaatgga aaatcaaaat ggtcgaattc atcagagctt aagataacaa attctagcat
     1861 agattttagg ctttacacac aagcgcgtct ctcggctatc tccctcactt accaggcatt
     1921 gtgtcttgga aaaggaaact atacggttaa tcttcatttc gctgaaatta tgttcaatga
     1981 aaagaacatg tatagcaatt tgggaagacg atacttcgac atatacgttc agggtaaacg
     2041 tgaagtgaaa gatttcaata tcgttgatga agcaaaaggt gtgggaaaag ctgttgttaa
     2101 gaaatttcct gtcatggtta caaatgggaa actggaaata agattgcagt gggctgggaa
     2161 aggaactcaa gctattcctg tgagaggtgt ctatggtcct ctcatatctg ctgtatcggt
     2221 agatccagat tttattccac caaaggaacc gggcactgga actggtggtg gaagttctct
     2281 ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc cccagatgct
     2341 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     2401 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
     2461 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     2521 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     2581 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     2641 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     2701 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     2761 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     2821 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     2881 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     2941 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     3001 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3061 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3121 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3181 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3241 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3301 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3361 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3421 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3481 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     3541 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     3601 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     3661 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     3721 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     3781 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     3841 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     3901 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
     3961 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
     4021 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     4081 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     4141 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4201 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4261 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4321 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     4381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6541 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6601 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6661 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6721 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6781 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6841 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6901 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6961 agggttttcc cagtcacgac gtt
//
