LOCUS       sequence-270261-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G60630 I8_pECIA14 should be used with bait vector 
            AT1G60630 I8_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  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 5897)
  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: AT1G60630 I8_pECIA14
FEATURES             Location/Qualifiers
     source          1..5897
                     /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             1193..1216
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2891..2914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2930..2947
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2970..2987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3020..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3098..3117
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3298..3314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3298..3314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3311..3333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3322..3338
                     /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(3346..3352,3353..3370,3371..3376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3391..3412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3529..3546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3700..4288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3780..3799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4459..5250,5251..5319))
                     /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     5082..5101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5320..5424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5492..5510
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5548..5570)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5670..5689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5883..5897,1..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 agcgatgttg aagctctact
      541 gagtttgaaa tcatccattg atccttcaaa ttcgattcca tggcgaggaa cagatccttg
      601 caactgggaa ggagtcaaga aatgcatgaa aggaagagtc tcaaagcttg ttcttgagaa
      661 tttgaacctc agtggctcac tcaacggcaa gagtttgaac caattggatc aacttagagt
      721 tcttagtttc aaaggaaact ctctctctgg ctcaatccct aacctatctg gtttagtcaa
      781 cctcaaatct ctttatctca acgataataa cttctccggt gagttccctg aatcactgac
      841 gtctctacac cgattgaaaa cggttgttct ttccaggaat cggttttccg gcaagattcc
      901 tagctcgttg ctccgactct cccgccttta cacgttttac gttcaagaca atctcttctc
      961 tggttccatt cctcctctca accaagcaac tctcagattc tttaatgtct cgaataatca
     1021 actctctggt catatcccac caacacaagc tctgaatcgg ttcaatgagt cgtcgtttac
     1081 tgacaacatt gctctctgcg gtgatcagat tcaaaactca tgcaatgaca ccactggaat
     1141 cacttcaact ccatcagcaa aaccagctat ccctgtagcc aaaacacgta gcctggaggt
     1201 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     1261 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     1321 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     1381 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     1441 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     1501 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     1561 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     1621 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     1681 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     1741 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     1801 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     1861 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     1921 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     1981 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2041 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2101 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2161 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2221 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2281 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     2341 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     2401 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     2461 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     2521 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     2581 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     2641 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     2701 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     2761 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     2821 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     2881 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     2941 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     3001 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     3061 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3121 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3181 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3241 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     3301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5701 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     5761 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5821 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5881 ttcccagtca cgacgtt
//
