LOCUS       sequence-270151-        6713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G21340 N4_pECIA14 should be used with bait vector 
            AT3G21340 N4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  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: AT3G21340 N4_pECIA14
FEATURES             Location/Qualifiers
     source          1..6713
                     /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             2009..2032
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3707..3730
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3746..3763
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3786..3803)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3836..3970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3860..3879)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3914..3933
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4114..4130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4114..4130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4127..4149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4138..4154
                     /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(4162..4168,4169..4186,4187..4192))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4207..4228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4345..4362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4596..4615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5275..6066,6067..6135))
                     /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     5898..5917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6136..6240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6308..6326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6364..6386)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6486..6505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6699..6713,1..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 gcgggtcatt
      541 gccaaatgag cctccttaca acgatccttc aaccggatta acatactcga cggacgatgg
      601 tttcgtgcag agtggcaaaa ctggaagaat ccagaaagcg ttcgagtcga tcttcagtaa
      661 accgtctttg aagcttagat acttcccgga cggattccga aactgctata ccttgaatgt
      721 cacgcaagac acaaactatc tgatcaaagc tgtatttgtg tatggtaact acgatggtct
      781 taacaatccc ccgagtttcg atctttacct tggtccgaat ctatgggtaa cggttgatat
      841 gaatggacgg accaatggta ctatccagga gattatccac aagaccatat ctaagtctct
      901 ccaggtctgt cttgttaaga caggaacaag ctcacctatg attaatacgt tagagctacg
      961 accacttaaa aacaatactt acaatactca gagtggctct ctgaagtatt tcttccgata
     1021 ttatttcagc ggttcaggcc aaaacatacg gtaccctgat gatgtcaatg atcgtaaatg
     1081 gtatccattc tttgatgcaa aagagtggac agagttaaca accaatctga atataaacag
     1141 ttctaatggt tatgcaccac cagaagttgt gatggcgtca gcctcaacgc ctataagtac
     1201 ttttggaaca tggaacttct catggttatt gccatcttcc acaacccaat tttatgtgta
     1261 catgcatttt gccgagattc aaactctacg gtccctcgat acccgagaat tcaaagtgac
     1321 gttgaatgga aaacttgctt atgaacgcta cagccctaaa acgttagcca ccgaaaccat
     1381 tttctattcg acaccacaac aatgtgaaga tgggacatgc ctcttggagt tgacgaaaac
     1441 acctaagtct actcttcctc ctctcatgaa cgctcttgag gttttcaccg tgatcgattt
     1501 tccacagatg gaaacaaatc cagatgatgt tgctgctatc aagagtatcc aaagcactta
     1561 tggattaagt aaaatcagct ggcaaggaga tccatgcgtt cctaaacagt ttttgtggga
     1621 gggtttaaac tgcaataatc tagataactc cacgccgcct attgtcactt ccttaaactt
     1681 atcgtcaagt catttaacgg ggatcatcgc gcaaggcatt cagaatctga cacacctaca
     1741 agaactagac ttgtcaaata acaatttgac gggaggaata cccgaatttc ttgctgacat
     1801 aaaatcactc ttagtaataa atttaagtgg gaacaatttt aatggctcta ttcctcaaat
     1861 ccttttacag aagaaaggac taaagctaat tcttgaagga aacgccaatc tgatttgtcc
     1921 ggatggatta tgtgtaaaca aagctggcaa tggtggtgcc aagaaaatga atgttgtaat
     1981 accgattgtt gcatcagttg cgtttgtgct ggaggtgctg ttccagggac caagcggcaa
     2041 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     2101 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     2161 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2221 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2281 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     2341 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     2401 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2461 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2521 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2581 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     2641 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     2701 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     2761 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     2821 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     2881 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     2941 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     3001 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     3061 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3121 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3181 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3241 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3301 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     3361 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3421 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3481 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3541 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3601 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     3661 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     3721 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3781 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3841 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3901 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3961 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4021 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     4081 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4141 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4201 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4261 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4321 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6241 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6301 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6361 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6421 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6481 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6541 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6601 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6661 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
