LOCUS       sequence-227239-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector SRF4 B5_pECIA14 should be used with bait vector SRF4 
            B5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  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 5957)
  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: SRF4 B5_pECIA14
FEATURES             Location/Qualifiers
     source          1..5957
                     /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             1253..1276
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2951..2974
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2990..3007
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3030..3047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3080..3214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3104..3123)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3158..3177
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3358..3374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3358..3374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3371..3393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3382..3398
                     /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(3406..3412,3413..3430,3431..3436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3451..3472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3589..3606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3760..4348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3840..3859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4519..5310,5311..5379))
                     /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     5142..5161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5380..5484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5552..5570
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5608..5630)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5730..5749
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5943..5957,1..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 agccaagacg tatctgctct
      541 taatgacgct tataagagca tgaactctcc atcaaaactc aaaggctggt cttcaagcgg
      601 aggcgatcct tgtggcgatt cgtgggatgg cattacttgc aaaggctctt ctgttactga
      661 aataaaagta tctggacgtg gactcagtgg atctttaggt taccagcttg gaaacttgaa
      721 atcgctcact taccttgatg taagcaagaa caatcttaac ggaaacttac cctaccagct
      781 tcctgacaag cttacttatc tagatggctc tgagaatgac ttcaacggta atgtgccata
      841 ttcggtgtct ctcatgaacg acctcagtta cctaaacctt ggccgtaaca acctcaatgg
      901 tgaactcagt gacatgtttc aaaagcttcc aaaacttgaa acaattgatc tttcttccaa
      961 tcaactcacg ggaaaactac cgcagagttt tgccaatcta acaggtctta aaacactgca
     1021 tttgcaagaa aaccaattca aaggctctat aaatgctctc agggacctcc ctcagattga
     1081 tgatgtaaac gtagcaaaca atcaatttac tggttggatc ccaaacgagt tgaagaatat
     1141 tggaaacctt gaaactggag gaaacaagtg gtcaagcggt agagctcctt caccacctcc
     1201 gggaacccgt cacatagaca gaaactcatc aggaggcggt ggaggaagca gcctggaggt
     1261 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     1321 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     1381 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     1441 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     1501 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     1561 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     1621 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     1681 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     1741 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     1801 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     1861 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     1921 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     1981 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2041 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2101 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2161 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2221 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2281 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2341 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     2401 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     2461 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     2521 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     2581 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     2641 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     2701 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     2761 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     2821 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     2881 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     2941 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     3001 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     3061 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     3121 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3181 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3241 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3301 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     3361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5521 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5581 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5641 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5701 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5761 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     5821 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5881 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5941 ttcccagtca cgacgtt
//
