LOCUS       sequence-270424-        6152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G37250 X020_pECIA14 should be used with bait vector 
            AT4G37250 X020_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  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 6152)
  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: AT4G37250 X020_pECIA14
FEATURES             Location/Qualifiers
     source          1..6152
                     /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             1448..1471
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3146..3169
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3185..3202
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3225..3242)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3275..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3299..3318)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3353..3372
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3553..3569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3553..3569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3566..3588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3577..3593
                     /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(3601..3607,3608..3625,3626..3631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3646..3667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3784..3801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3955..4543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4035..4054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4714..5505,5506..5574))
                     /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     5337..5356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5575..5679)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5747..5765
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5803..5825)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5925..5944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6138..6152,1..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 tctgctttaa actctgatgg
      541 gttggttttg atgaagttta aatcctctgt tcttgttgat cctctgtctt tattacagac
      601 atggaattac aaacacgaaa gtccatgttc ttggagagga atctcttgta ataacgattc
      661 taaagtctta actttgtctc ttcccaattc tcagcttctt ggttcgattc cttctgattt
      721 gggttctctc ttaacacttc aaagtctcga tctttccaat aattccttca atgggccatt
      781 accggtttct ttcttcaacg ctagagagct tcgtttcctt gatctctcta gtaacatgat
      841 ctccggcgaa atcccttccg caattggtga tcttcacaat cttctaaccc taaatctctc
      901 tgataatgcc ttggcgggaa agttacctac aaatttggcg tcgttgagaa acttaacggt
      961 ggtttctttg gagaataatt acttttccgg cgagattccc ggcggttgga gagttgttga
     1021 gttcttagac ctgtcgtcga atctgatcaa tggctcgctt cctccggatt ttggcggata
     1081 tagtctccag tacttaaatg tctcgtttaa ccaaatctcc ggcgaaattc caccggagat
     1141 cggagtcaat ttcccaagaa acgtcaccgt tgatctctcc ttcaacaatt taaccggtcc
     1201 aatccctgat tctccggttt tcttaaacca ggaatcaaat ttcttttcag gaaacccggg
     1261 tctatgcggt gaaccgaccc gaaacccgtg tcttatccct tcctcgccgt ctatcgtctc
     1321 cgaagccgac gtaccgacgt ctacaccagc aatcgccgca ataccgaaca caatcggttc
     1381 aaacccggtt accgatccaa attcacaaca aaccgaccca aatccccgaa ccgggttacg
     1441 acccggactg gaggtgctgt tccagggacc aagcggcaat cccggcggcg atctggcccc
     1501 ccagatgctg cgcgagctgc aggagaccaa tgccgccctg caggacgtgc gcgagctgct
     1561 gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc gtgatggagt gcgacgcctg
     1621 cggcatgcag ccctctagag gctccggctc catcatccca gttgaggagg agaacccgga
     1681 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     1741 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     1801 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     1861 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     1921 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     1981 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     2041 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     2101 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     2161 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     2221 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     2281 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     2341 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     2401 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     2461 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     2521 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     2581 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     2641 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     2701 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     2761 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     2821 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     2881 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     2941 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     3001 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     3061 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggttc
     3121 tagaggaagc acctccggcg ataacgatta caaggatgat gatgataagg ccagcggcac
     3181 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3241 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3301 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3361 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3421 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3481 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     3541 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3601 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3661 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3721 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3781 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3841 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4381 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4801 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4921 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5161 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5221 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5401 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5581 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5641 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5701 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5761 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5821 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5881 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5941 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6001 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6061 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6121 ggtaacgcca gggttttccc agtcacgacg tt
//
