LOCUS       sequence-275961-        7463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BRL3 C7_pECIA14 should be used with bait vector BRL3 
            C7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7463)
  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 7463)
  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: BRL3 C7_pECIA14
FEATURES             Location/Qualifiers
     source          1..7463
                     /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             2759..2782
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4457..4480
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4536..4553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4586..4720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4610..4629)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4664..4683
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4864..4880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4864..4880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4877..4899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4888..4904
                     /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(4912..4918,4919..4936,4937..4942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4957..4978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5095..5112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5266..5854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5346..5365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6025..6816,6817..6885))
                     /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     6648..6667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6886..6990)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7058..7076
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7114..7136)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7236..7255
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7449..7463,1..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 agacggctac tcagcgacga
      541 cgttaacgac acagctctgc taacggcgtt taagcaaacc tctatcaagt cggatcctac
      601 caacttttta ggtaactgga gatacgggtc gggtcgtgac ccatgtactt ggcgaggcgt
      661 ctcttgttct agcgatggta gagtcattgg acttgatcta cgaaacggtg gtcttaccgg
      721 aaccctaaat ctcaataatc tcacggcgtt gtctaatctc cggagtcttt atttgcaagg
      781 aaacaacttc tcctccggag attcttcttc ttcttcagga tgttctctcg aggttctgga
      841 tttatcttca aactcgctta cggactcttc aatagtggat tacgttttct ccacctgttt
      901 aaatctggtt tctgtcaatt tctcgcacaa caagctcgcc gggaaattga agtcttctcc
      961 ttcggcgagt aacaagagaa tcacaaccgt tgatctctcg aacaatcgct tctccgatga
     1021 gattccggag actttcatcg ccgattttcc gaattctctg aaacacctcg acttatccgg
     1081 caacaatgtc accggcgact tctctcgtct cagctttggt ctctgtgaga acctcaccgt
     1141 attcagctta tcacagaaca gcatctccgg cgacagattt ccggttagtc tctcaaattg
     1201 taagctcctc gaaacgttaa acctctctcg gaacagtctc atcggaaaaa tccccggaga
     1261 tgactattgg ggaaattttc agaacctgag gcagctctct cttgcacaca acctttactc
     1321 cggtgagatt ccaccggagc tttctctgct ctgtcgaact ctagaggttc tcgatctctc
     1381 cggcaacagt ctcaccggtc agcttccgca atcattcacc tcttgtggct cattacaaag
     1441 cctaaatctt ggaaacaaca agctctccgg tgatttctta agcaccgtcg tgagtaaact
     1501 ctctagaatc actaatcttt acttaccatt caacaacatc tcaggctctg ttccgatttc
     1561 gctcaccaac tgtagtaatc tgcgagttct tgatctgagc tcaaacgagt tcactggaga
     1621 agtaccatct ggtttctgct ctcttcaaag ctcttcggtt ttggaaaagt tgcttattgc
     1681 caacaattac ctctcaggaa ctgtccctgt agagcttggt aaatgtaaga gcttgaagac
     1741 tatagatctt agcttcaatg ctctcactgg tctgattcct aaggagatat ggacattgcc
     1801 taagctctcc gacttagtta tgtgggcgaa caatctcact ggtggaatcc cggagagcat
     1861 ttgtgttgat ggaggaaact tggaaacgtt aatcttaaac aacaatctct taaccggttc
     1921 tcttccagaa tcaatctcca aatgcaccaa catgctttgg atctcccttt ctagtaacct
     1981 gcttaccggt gaaatccctg taggaatcgg aaagcttgag aagttagcga ttctgcaact
     2041 cgggaacaat tctttaactg ggaacattcc aagtgagctt ggaaactgca agaaccttat
     2101 ctggcttgat cttaacagca ataatctaac tgggaatctc ccaggtgaac ttgcaagtca
     2161 ggccggactg gtaatgcccg gaagcgtctc tggtaaacag tttgcgtttg tgaggaatga
     2221 aggtgggacg gattgcagag gtgcaggggg tttggttgag tttgaaggaa ttcgtgctga
     2281 gcggttagag cattttccaa tggttcattc atgtcccaag actcggatat actcaggcat
     2341 gaccatgtac atgttctcca gtaacgggag catgatctac ttggaccttt cctacaatgc
     2401 ggtttcaggt tctattcctt tgggttatgg tgcaatggga tatcttcagg tcttgaattt
     2461 gggacataac ttgttaactg gaaccatacc ggatagcttc ggaggattga aagcaattgg
     2521 agttcttgat ctatctcaca atgatctcca aggattctta cctggatcac taggaggact
     2581 ctcttttcta agcgacttag acgtctctaa caacaatcta accggaccaa tcccattcgg
     2641 aggccaactt acgacgttcc cactcacaag atacgccaac aactctggcc tctgcggggt
     2701 tcctctacca ccttgtagct ctggttcccg gcccacgaga tctcatgctc accctaaact
     2761 ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc cccagatgct
     2821 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     2881 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
     2941 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     3001 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     3061 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     3121 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     3181 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     3241 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     3301 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     3361 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     3421 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     3481 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3541 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3601 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3661 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3721 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3781 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3841 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3901 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3961 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     4021 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     4081 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     4141 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     4201 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     4261 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     4321 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     4381 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
     4441 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
     4501 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     4561 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     4621 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4681 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4741 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4801 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     4861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5461 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5701 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7021 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7081 tcgtctcgcg cgttttggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7141 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7201 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7261 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7321 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7381 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7441 agggttttcc cagtcacgac gtt
//
