LOCUS       sequence-270225-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G45780 I3_pECIA14 should be used with bait vector 
            AT5G45780 I3_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  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 5834)
  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: AT5G45780 I3_pECIA14
FEATURES             Location/Qualifiers
     source          1..5834
                     /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             1130..1153
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2828..2851
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2867..2884
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2907..2924)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2957..3091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3035..3054
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3235..3251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3235..3251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3248..3270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3259..3275
                     /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(3283..3289,3290..3307,3308..3313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3328..3349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3466..3483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3637..4225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3717..3736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4396..5187,5188..5256))
                     /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     5019..5038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5257..5361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5429..5447
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5485..5507)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5607..5626
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5820..5834,1..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 tctcccaagg gtgttaacta
      541 tgaagtggct gcgttaatgt cagtgaagaa caagatgaaa gatgagaaag aggttttgtc
      601 tggttgggat attaactctg ttgatccttg tacttggaac atggttggtt gttcttctga
      661 aggttttgtg gtttctctag agatggctag taaaggatta tcagggatac tatctactag
      721 tattggggaa ttaactcatc ttcatacttt gttacttcag aataatcagt taactggtcc
      781 gattccttct gagttaggcc aactctctga gcttgaaacg cttgatttat cggggaatcg
      841 gtttagtggt gaaatcccag cttctttagg gttcttaact cacttaaact acttgcggct
      901 tagcaggaat cttttatctg ggcaagtccc tcacctcgtc gctggcctct caggtctttc
      961 tttcttggat ctatctttca acaatctaag cggaccaact ccgaatatat cagcaaaaga
     1021 ttacaggatt gtaggaaatg catttctttg tggtccagct tcccaagagc tttgctcaga
     1081 tgctacacct gtgagaaatg cgacgggttt gtctgaaaag gacaatagcc tggaggtgct
     1141 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     1201 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     1261 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     1321 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     1381 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     1441 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     1501 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     1561 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     1621 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     1681 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     1741 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     1801 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     1861 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     1921 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     1981 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     2041 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     2101 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     2161 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     2221 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     2281 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     2341 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     2401 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     2461 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     2521 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     2581 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     2641 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     2701 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     2761 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     2821 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     2881 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     2941 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3001 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3061 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3121 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3181 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     3241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5821 ccagtcacga cgtt
//
