LOCUS       sequence-270168-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G16900 O2_pECIA14 should be used with bait vector 
            AT5G16900 O2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  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 6674)
  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: AT5G16900 O2_pECIA14
FEATURES             Location/Qualifiers
     source          1..6674
                     /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             1970..1993
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3668..3691
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3707..3724
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3747..3764)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3797..3931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3821..3840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3875..3894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4075..4091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4075..4091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4088..4110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4099..4115
                     /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(4123..4129,4130..4147,4148..4153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4168..4189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4306..4323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4477..5065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4557..4576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5236..6027,6028..6096))
                     /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     5859..5878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6269..6287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6325..6347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6447..6466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6660..6674,1..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 agtttggatt gcgggttgcc
      541 gagtaacgag cctccatata ttgaaccggt aaccggatta gtattctcat cggatgcgga
      601 tcacatccct agtggaataa gcggtagaat ccagaagaat ctggaagcag tacacataaa
      661 gccgtatttg tttctgagat actttccaga tgggctacga aattgctata ctctagatgt
      721 ccttcagaac cgaagataca tgatcaaagc tgtgtttgtg tatggaaatt acgatggtta
      781 taatgattat ccgagttttg atctgtatct aggtcctaat aagtgggtaa gagttgactt
      841 ggaaggaaag gtcaatggca gtgttgaaga gattattcac ataccaagtt caaactctct
      901 gcagatttgt cttgtcaaga caggtaattc tttgccgttc atatccgccc tggaactgag
      961 gcttttgaga aacgatactt atgttgtcca agacgtttcc ttgaagcatt tatttcggag
     1021 gtactataga cagtcagatc ggttaatccg gtatcctgac gatgtctatg atcgtgtttg
     1081 gtccccattc ttcttaccgg aatggacgca gataactacc agtttagatg tgaataactc
     1141 taacaactac gagccaccaa aagcagcatt aacatcagct gccacacctg gagacaatgg
     1201 aacgcgattg accattattt ggactttgga taaccctgat gagcaaattc acctgtatgt
     1261 tcactttgcc gagcttgaac ctgttgggga aaacactgac gaggccttga gaactttatt
     1321 tacaaggacc ttttactttg tggtgaatgg aaagattagt tatgatgaga gtattactcc
     1381 tttagacttg gctgtaagta ctgtagagac tgtggtgaac aaatgcgatg gcggtaactg
     1441 cagtttgcag ctggtacgca gcgaagcgtc tccgggagta cgtgtacctc tggttaatgc
     1501 tatggaggca ttcactgcaa tcaagtttcc acattctgaa acaaatcctg atgatgttat
     1561 atctatcaag gttatccaag ctacctatga actgagcaga gtcgactggc aaggagatcc
     1621 gtgtcttccc cagcagtttt tgtggactgg cctaaactgc agctacatga acatgtctac
     1681 gtcaccaaga atcatttcct tagacctgtc ctcacataaa ttaactggga aaatagtgcc
     1741 agatatacaa aatctgaccc agctacagaa actggacttg tcgaataaca agttgaccgg
     1801 aggtgtgcct gagtttctag ccaacatgaa gtcactgttg ttcataaacc taagcaataa
     1861 caatcttgtt ggttccattc ctcaagcgct tcttgaccga aaaaatctga agttggagtt
     1921 tgaaggaaat ccaaagcttt gcgccacagg tccgtgtaac agttcaagtc tggaggtgct
     1981 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     2041 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     2101 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     2161 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     2221 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     2281 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     2341 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     2401 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     2461 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     2521 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     2581 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     2641 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     2701 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     2761 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     2821 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     2881 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     2941 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     3001 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     3061 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     3121 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     3181 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     3241 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     3301 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     3361 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     3421 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     3481 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     3541 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     3601 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     3661 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     3721 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     3781 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3841 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3901 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3961 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4021 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     4081 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4141 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4201 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6301 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6361 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6421 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6481 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6541 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6601 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6661 ccagtcacga cgtt
//
