LOCUS       sequence-238710-        4898 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector FEI2 X041_pECIA2 should be used with prey vector FEI2 
            X041_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4898)
  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 4898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: FEI2 X041_pECIA2
FEATURES             Location/Qualifiers
     source          1..4898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1596..1619
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2325..2366
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2376..2393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2416..2433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2466..2600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2490..2509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2544..2563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2744..2760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2757..2779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2768..2784
                     /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(2792..2798,2799..2816,2817..2822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2837..2858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2975..2992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3146..3734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3226..3245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3905..4696,4697..4765))
                     /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     4528..4547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4766..4870)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggaatga agcaattagt cctgatggtg aggcgctttt gagttttaga aatggagttt
     1021 tggcttctga tggtgttatt ggtctgtgga gaccagagga tcctgatcca tgtaactgga
     1081 aaggagtgac ctgtgatgca aaaacaaaaa gagttatagc cttgagtctt acttatcaca
     1141 aattaagggg acctttaccc cctgagcttg gaaagctgga tcagttaagg cttttaatgc
     1201 ttcacaacaa tgcgttatat caatcaatac ctgcctcatt gggaaattgc acggcattag
     1261 agggaatata cttgcagaat aattacatca ctgggacaat cccaagtgaa ataggaaacc
     1321 tgtccgggct gaaaaacctg gacctctcaa acaacaatct caatggagct atccctgctt
     1381 cacttgggca gttgaaaagg cttactaaat tcaatgtctc aaacaatttc ctggtgggaa
     1441 aaataccttc tgacggccta ctcgctagac tctcgaggga ctccttcaac ggaaatcgta
     1501 acttgtgtgg aaaacaaatc gatattgtgt gcaatgacag tggaaattct acagcttctg
     1561 ggtctccaac aggccaagga ggtaataacc ctaaactgga ggtgctgttc cagggaccag
     1621 ggagcgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt
     1681 cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg
     1741 tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg
     1801 tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca
     1861 cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt
     1921 acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag
     1981 ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga
     2041 ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg
     2101 tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg
     2161 actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc
     2221 aggggaacgt cttctcatgc tccgtgatgc atgagggtct gcacaaccac tacacgcaga
     2281 agagcctctc cctgtctccg ggtaaatcta gagggccctt cgaaggtaag cctatcccta
     2341 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     2401 aaacccgctg atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga
     2461 tcgaggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2581 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2641 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     2701 atgatcagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     2761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2941 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4861 gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
