LOCUS       sequence-227455-        5027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector SRF8 B8_pECIA2 should be used with prey vector SRF8 
            B8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  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 5027)
  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: SRF8 B8_pECIA2
FEATURES             Location/Qualifiers
     source          1..5027
                     /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             1280..1303
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2009..2050
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2060..2077
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2100..2117)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2150..2284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2174..2193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2228..2247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2428..2444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2428..2444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2441..2463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2452..2468
                     /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(2476..2482,2483..2500,2501..2506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2521..2542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2659..2676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2830..3418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2910..2929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3589..4380,4381..4449))
                     /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     4212..4231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4450..4554)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4622..4640
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4678..4700)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4800..4819
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5013..5027,1..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 gatccatctg atgttcaagc
      541 ccttcaagtt ttgtatactt cactaaatag tccttcacag ctaaccaatt ggaaaaatgg
      601 tggtggtgat ccatgtggag agtcatggaa agggattact tgtgagggct ctgcagttgt
      661 cactatagat atatcggatt taggagtctc tggcactctt ggatacttgc tctcagacct
      721 taagtcactc agaaaattgg atgttagtgg taacagcatc catgatacac ttccatatca
      781 gttgccaccg aatctcacaa gcctaaatct tgcgagaaac aacctaagtg gtaaccttcc
      841 atactccata tccgccatgg gttctctttc gtatatgaat gtaagtggta attcacttac
      901 aatgtcaata ggagatatct ttgctgatca taagtcactg gcaaccttgg acctttctca
      961 taacaacttc agtggcgatc ttcctagttc cttaagcaca gtgtccacac tttccgttct
     1021 ttatgttcag aacaatcaat tgacaggttc catcgatgtt ctctcggggt tgcctttgaa
     1081 aactttaaat gttgcaaaca atcatttcaa tggatccata cctaaggagc ttagttctat
     1141 tcagacctta atatatgatg gtaattcctt tgataacgtc cctgcctctc ctcagccaga
     1201 aagacctggc aaaaaagaga caccttctgg ttctaagaaa cccaaaattg gttctgaaga
     1261 gaaatcttca gactctggcc tggaggtgct gttccaggga ccagggagcg acaaaactca
     1321 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     1381 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     1441 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     1501 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     1561 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     1621 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     1681 agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag
     1741 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     1801 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     1861 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     1921 atgctccgtg atgcatgagg gtctgcacaa ccactacacg cagaagagcc tctccctgtc
     1981 tccgggtaaa tctagagggc ccttcgaagg taagcctatc cctaaccctc tcctcggtct
     2041 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2101 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     2161 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2221 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2281 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2341 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     2401 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2461 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2521 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2641 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2701 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2761 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3241 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3601 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3661 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3721 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3781 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3841 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3901 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3961 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4021 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4081 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4141 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4201 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4261 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4321 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4381 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4441 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4501 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4561 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4621 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4681 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4741 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4801 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4861 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     4921 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     4981 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
