LOCUS       sequence-273952-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G03770 O12_pECIA14 should be used with bait vector 
            AT3G03770 O12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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: AT3G03770 O12_pECIA14
FEATURES             Location/Qualifiers
     source          1..6293
                     /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             1589..1612
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3287..3310
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3326..3343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3366..3383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3416..3550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3440..3459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3494..3513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3694..3710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3694..3710)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3707..3729)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3718..3734
                     /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(3742..3748,3749..3766,3767..3772))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3787..3808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4855..5646,5647..5715))
                     /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     5478..5497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5716..5820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5888..5906
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5944..5966)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6066..6085
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6279..6293,1..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 tctcaaaccc tcttgaggct
      541 gcaacagctt ctctactatc ccaaagtgtt aaacagctgg aacaactata cagatttctg
      601 caattctgag ccaagccctt cgctaaccgt tgtgtgttat gaagatagtg taacgcagct
      661 tcacattata ggagacaatg gaactcatat gttaccaaag agtttctcaa taaactcgtt
      721 tgttactact cttgttaagc tccctgatgt taaagttctc acgtttgtgt cccttgggtt
      781 atggggttgg ttacctcaga aaatcaaccg tttatcctct ttggagattc tcaatgtgag
      841 ttccaatttc ctctttggac caatccctca tgaactctcg tcgcttgcta ctcttcagac
      901 gcttatactc gacgagaata tgttttctgg tgagttaccg gattggattg attctttgcc
      961 gagtttagct gtgttgagct tgagaaagaa cgtgcttaat gggtcattgc cttcttcttt
     1021 gagtagcttg tcaggtctta gagttcttgc tttagctaat aaccggttta atggagcttt
     1081 gcctgattta agccatttga caaaccttca agtgcttgat cttgaaggaa actcttttgg
     1141 accgttgttt cctcggttaa gtaacaagct ggttactctt atactcagca agaacaagtt
     1201 tagatctgca gtttcagctg aagaagtaag ttctctgtat cagcttcaac atttggatct
     1261 ttcctacaat acattcgttg gtccatttcc tacttccttg atgtctcttc ccgcgataac
     1321 ttacttgaac atttcacaca acaaactcac cgggcggctt tcagcaaatc tttcttgcaa
     1381 ctcacagctg atgtttgtag atatgtcatc aaatcttctt accggaagtt tgcctacttg
     1441 ccttaaaccg agttccggca ctagcagaga tgttgtctat gcaagcaact gtttggctac
     1501 tacgaatgaa gatcaacgtc cagtttcttt ttgtagtaat gaagctcttg ctgttggaat
     1561 cctaccacag aggaggaaca aagtttccct ggaggtgctg ttccagggac caagcggcaa
     1621 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     1681 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     1741 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     1801 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     1861 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     1921 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     1981 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2041 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2101 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2161 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     2221 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     2281 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     2341 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     2401 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     2461 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     2521 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     2581 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     2641 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     2701 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     2761 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     2821 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     2881 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     2941 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3001 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3061 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3121 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3181 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     3241 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     3301 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3361 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3421 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3481 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3541 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3601 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     3661 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3721 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3781 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3841 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3901 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5761 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5821 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5881 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5941 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6001 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6061 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6121 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6181 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6241 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
