LOCUS       sequence-270173-        6803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G48740 O3_pECIA14 should be used with bait vector 
            AT5G48740 O3_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6803)
  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 6803)
  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: AT5G48740 O3_pECIA14
FEATURES             Location/Qualifiers
     source          1..6803
                     /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             2099..2122
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3797..3820
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3836..3853
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3876..3893)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3926..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4004..4023
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4204..4220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4204..4220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4217..4239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4228..4244
                     /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(4252..4258,4259..4276,4277..4282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4297..4318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4435..4452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4606..5194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4686..4705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5365..6156,6157..6225))
                     /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     5988..6007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6226..6330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6398..6416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6454..6476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6576..6595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6789..6803,1..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 agtccagatg gtttcttgag
      541 cttatcatgt ggcggctcaa gttacaccgc tgcttataac atctcatggg tttcagacaa
      601 cgattatatc gaaacaggta acactaccac ggttacttac gccgagggga attctacttc
      661 cagtgttccc atcagattat tcccggatcc tcaaggtcgc caatgttaca agttacctgt
      721 gaggaaagac ttgtcttcag ttctgattcg tgctacattc gtttatagaa actatgatag
      781 ccagaatagt cctccagcat ttcatgtctc tcttggaagg agaatcacta gtactgttga
      841 tcttagaacc aatgatcctt ggattgaaga gttagtttgg ccagtcaaca atgattctct
      901 tttgctttgc ttgcttgcgg ttaaaggaag aggtattccg gttatctctt ctctagaagt
      961 acggccactt cctttaggtt cttacaaata cagcctggag ggttcaccag atataattct
     1021 caggagaagt taccgaatca atagcggata cacaaatggg actatccggt atccttcaga
     1081 tccatttgat cgaatatggg atccggatca aagttattca ccgtttcacg cctcttggag
     1141 cttcaatgga ctcacaaaat tgaattcctt caatattact gagaatccac cagcttctgt
     1201 tcttaagaca gcaagaattc ttgctcgcaa agaaagtctt tcatatactc tttctctaca
     1261 tacaccaggg gactactaca ttatcctcta ctttgctggg atactctcac tctccccttc
     1321 ttttagtgtg acgatcaatg atgaagtcaa acaatctgat tacaccgtaa ctagctcaga
     1381 agctggtacc ttgtacttta cgcaaaaagg aatcagtaag ttaaacatca ccctcagaaa
     1441 aatcaagttc aatcctcaag tgagtgctct tgaggtatat gaaatcctcc aaatccctcc
     1501 tgaagcttct tctaccacag tttcggcact taaagtcata gagcaattta ccgggcaaga
     1561 ccttggatgg caagacgacc catgcacacc gcttccttgg aaccacatcg aatgtgaagg
     1621 aaaccgtgtt acatcattgt tcctttcaaa aatcaacctg aggtccatta gtccaacatt
     1681 cggagacttg ctcgatctca aaacactcga cttgcataat acatcactta caggagcaat
     1741 acagaatgta ggcagcctta aggatctcca gaaactgaat ttgagcttca atcagttaga
     1801 atcttttggc tccgagctgg aagatctggt taaccttgaa gttctggacc tgcaaaacaa
     1861 cagcttacaa ggatcagttc ctgaaacatt aggaaagctg aagaaacttc gtctcctgaa
     1921 tctggaaaac aataacttag taggtccctt gccacagtct cttaacataa cgggtctgga
     1981 agtcaggata acgggaaatc catgcctttc gttctcttca atatcatgca acaatgtgtc
     2041 ttcaacaatt gacacaccac aagtcactat ccctatcaat aagaaacaaa gaaaacagct
     2101 ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc cccagatgct
     2161 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     2221 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
     2281 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     2341 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     2401 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     2461 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     2521 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     2581 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     2641 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     2701 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     2761 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     2821 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     2881 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     2941 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3001 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3061 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3121 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3181 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3241 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3301 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     3361 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     3421 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     3481 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     3541 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     3601 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     3661 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     3721 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
     3781 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
     3841 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     3901 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     3961 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4021 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4081 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4141 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     4201 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4261 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4321 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4381 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4441 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4501 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5041 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5281 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5401 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5461 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5521 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5581 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5641 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5701 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5761 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5821 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5881 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5941 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6001 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6061 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6361 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6421 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6481 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6541 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6601 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6661 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6721 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6781 agggttttcc cagtcacgac gtt
//
