LOCUS       sequence-276501-        7343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G35710 X075_pECIA14 should be used with bait vector 
            AT1G35710 X075_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7343)
  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 7343)
  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: AT1G35710 X075_pECIA14
FEATURES             Location/Qualifiers
     source          1..7343
                     /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             2639..2662
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4337..4360
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4376..4393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4416..4433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4466..4600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4490..4509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4544..4563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4744..4760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4744..4760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4757..4779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4768..4784
                     /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(4792..4798,4799..4816,4817..4822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4837..4858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4975..4992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5226..5245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5905..6696,6697..6765))
                     /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     6528..6547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6766..6870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6938..6956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6994..7016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7116..7135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7329..7343,1..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 tctgcaacta tcgcagaagc
      541 aaatgctctt ctgaaatgga aatccacttt cacgaactca tcaaaactgt cttcatgggt
      601 ccatgatgca aacaccaata ctagcttttc ctgcaccagt tggtatggtg tttcttgcaa
      661 ctcacgaggg agtatcgaag agttaaacct cactaacact ggtatagaag gtactttcca
      721 agatttcccc ttcatttctc tctcaaacct tgcttacgta gatcttagca tgaacctttt
      781 atctggaacc atccctcctc aattcggaaa cctctctaaa ctcatctact ttgacctctc
      841 cacaaatcac ttaaccggag aaatctctcc ttccttagga aatctcaaga acttgacggt
      901 actttatctt caccaaaact atctaactag tgtcattccc tcggaactag gcaacatgga
      961 atccatgaca gatttagcgc taagtcagaa caaacttacc ggttcaattc cttcttcctt
     1021 aggaaatctc aagaacttga tggttcttta tctttacgag aactatctaa ctggtgtcat
     1081 tcccccrgaa mtaggyaaca tggaatccat gacarattta gckytaagtc agaacaaact
     1141 tactggttca attccttctt ccttaggtaa tctcaagaac ttgacgcttc tttctctttt
     1201 ccagaactat ctaactggtg gcattccccc gaaactaggc aacatagaat ccatgatcga
     1261 tttagagttg agtaacaaca aacttaccgg ttcaattcct tcttccttag gaaatctcaa
     1321 gaatttgact attctttatc tttacgagaa ctatttaact ggtgtcattc ccccggaact
     1381 aggcaacatg gaatcgatga ttgatttaca gttgaataac aacaaactta ccggttcaat
     1441 tccttcttcc ttcggaaatc tcaagaactt gacgtatctt tatctttacc tgaattattt
     1501 aactggtgtc attccccagg agctaggcaa catggaatcc atgatcaatt tagatttaag
     1561 tcagaacaaa cttacaggtt ctgttcctga ttcctttggt aactttacca agctggaaag
     1621 tttgtacctt cgtgttaacc acctctctgg tgcgatcccg ccaggagtag caaactcttc
     1681 acatctaact accttgatac ttgacaccaa caacttcact ggtttctttc ctgaaactgt
     1741 ttgcaaaggt aggaagcttc aaaatatctc attggattac aatcacctcg aaggtcctat
     1801 cccgaaaagc ttgagagatt gcaagagcct gatcagagca agatttctag gaaacaaatt
     1861 cactggcgat atttttgaag cttttgggat ttacccagac cttaacttca ttgatttcag
     1921 ccacaacaaa ttccacggtg agatttctag caactgggag aagagtccaa agctaggtgc
     1981 cttgatcatg tcaaacaaca acatcacggg tgctatccca acagagattt ggaacatgac
     2041 acaactagtt gagctggatc tatctaccaa caacctcttt ggtgaacttc cagaagcaat
     2101 tggaaatctc acgaatttgt cgaggctacg acttaatggg aatcagttat ctggaagagt
     2161 tcctgcaggt ttaagtttct taaccaatct tgagtctctt gacttatcct caaacaattt
     2221 cagctcagag atcccacaaa cctttgactc ctttttaaag cttcatgaca tgaacctgag
     2281 cagaaacaaa ttcgacggaa gcattccccg actatcaaag ctgactcagt taacgcaact
     2341 tgatctcagc cacaaccagc tcgacggaga aatcccatca caactaagct ccctgcaaag
     2401 cctcgacaag ctcgacctct cacacaacaa tctctcaggt ctcattccaa caactttcga
     2461 gggcatgata gcgctgacga acgtcgatat atcaaacaat aaactcgaag gtccgcttcc
     2521 agatactcca acgtttcgaa aagcaacagc agatgcattg gaggaaaaca taggcttatg
     2581 cagtaatatt cctaaacaaa gattgaaacc atgccgtgag ttgaagaaac cgaagaaact
     2641 ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc cccagatgct
     2701 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     2761 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
     2821 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     2881 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     2941 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     3001 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     3061 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     3121 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     3181 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     3241 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     3301 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     3361 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3421 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3481 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3541 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3601 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3661 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3721 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3781 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3841 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     3901 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     3961 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     4021 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     4081 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     4141 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     4201 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     4261 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
     4321 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
     4381 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     4441 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     4501 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4561 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4621 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4681 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     4741 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6961 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7021 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7081 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7141 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7201 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7261 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7321 agggttttcc cagtcacgac gtt
//
