LOCUS       sequence-298778-        6692 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector EMS1 X044_pECIA2 should be used with prey vector EMS1 
            X044_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  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 6692)
  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: EMS1 X044_pECIA2
FEATURES             Location/Qualifiers
     source          1..6692
                     /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             2945..2968
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3674..3715
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3725..3742
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3765..3782)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3815..3949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3839..3858)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3893..3912
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4093..4109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4093..4109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4106..4128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4117..4133
                     /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(4141..4147,4148..4165,4166..4171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4186..4207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4324..4341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4575..4594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5254..6045,6046..6114))
                     /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     5877..5896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6115..6219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6287..6305
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6343..6365)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6465..6484
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6678..6692,1..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 tccgccattg ttgatcttag
      541 ctcggaaaca acttctctca tctccttcaa gcgctcacta gaaaacccat ctctcctttc
      601 ttcatggaac gtttcgtctt ccgcgtctca ctgcgattgg gttggcgtta cgtgtctgct
      661 tggacgagtc aactcgctct ccttaccgtc tttgtctctg agaggccaga ttccgaagga
      721 aatttcgagt ttgaagaacc ttagagagct ttgtcttgcc ggaaatcaat tctccgggaa
      781 gattccgcct gagatttgga atctaaaaca tctccaaacc cttgacctct caggaaattc
      841 tctcaccgga cttctcccga ggctactcag tgagttacct cagcttctct acctcgacct
      901 tagtgacaac cacttctccg gttcacttcc tccatcgttc ttcatcagct tacctgcact
      961 gtcttctctt gacgtctcca acaactctct ctccggcgaa atccctcctg aaatcgggaa
     1021 actgagtaac ctctccaatc tttacatggg actcaactca ttttccggtc aaattccgtc
     1081 ggagattggt aacatctcgt tgctgaagaa ctttgctgca ccttcttgct tcttcaatgg
     1141 accgttacca aaggagattt cgaaactgaa acacttagca aagctcgacc tttcttataa
     1201 tccactcaag tgttccattc ccaaatcgtt tggagagttg cataatctga gtatactaaa
     1261 tcttgtctcc gcagagctta ttggattgat cccaccagag cttggtaatt gtaaaagcct
     1321 caagtcactg atgctgtctt tcaattctct ctctggacct ttgccattgg agctttcaga
     1381 gattcctctg ttgacgtttt ctgctgaaag gaatcagctt tcgggttcat tgccttcatg
     1441 gatggggaaa tggaaggtac tggactcact tctgttagct aacaatcggt tttcaggtga
     1501 aatccctcat gagattgagg attgtccaat gttgaaacat ctcagtctag ctagcaactt
     1561 gttaagtggt tcgattccta gggagctctg tggttcaggt tcgttggagg cgattgatct
     1621 ctctgggaat ttgctttctg gaacaattga ggaggtcttt gatggatgtt cctctcttgg
     1681 agagttgtta cttactaata atcagatcaa tggctctatt cctgaggatc tttggaagct
     1741 tccattgatg gcacttgact tggattccaa caacttcaca ggagaaatcc caaagagtct
     1801 ctggaaatca acgaacttga tggagtttac agcttcttat aatcgtctag aaggttattt
     1861 gcctgcagag attggtaatg cagcttcact gaagcggttg gttctcagtg ataatcagtt
     1921 aacaggtgaa atcccaagag agattggaaa actcacatct ttgtctgtct tgaatctgaa
     1981 tgcaaacatg tttcaaggga agataccagt tgagctcggt gattgtactt ctttgactac
     2041 attggatcta ggcagcaaca atcttcaagg acagattcca gacaaaatca ctgctctcgc
     2101 tcagcttcag tgcctggttc tctcttacaa caatctctcc gggtcaattc catcaaaacc
     2161 ctctgcttat ttccatcaga ttgagatgcc tgatctcagc tttcttcagc accatgggat
     2221 atttgatctc tcctacaata gattgtctgg ccctatacca gaagaacttg gcgagtgttt
     2281 ggtattggta gagatttcac tgagcaacaa ccatctctct ggagagattc cagcttcact
     2341 ctctcgtttg accaacctca caatcctgga tctatctgga aatgcactga ctggatctat
     2401 tccaaaggag atgggcaact ctcttaagtt gcagggatta aacttggcga ataatcagct
     2461 caatggccat attccagaaa gctttggctt gttaggtagc ttggtgaagc taaacttgac
     2521 taagaacaag cttgatggac cggttcctgc ttctcttggg aacttgaaag agcttacaca
     2581 catggacttg agctttaaca acctaagtgg cgagctttca tcagaactgt ctacaatgga
     2641 aaaacttgtc ggcctctaca ttgagcagaa taagttcacc ggggaaatac cttcagaact
     2701 tggtaatctt acgcagctgg agtaccttga tgtttctgag aacttgttat ctggagagat
     2761 accaacaaag atctgtggat tgcccaatct ggagttcttg aatctagcca agaacaatct
     2821 gcgaggagag gtgccgagtg atggtgtttg tcaggatcct tccaaggcat tgctttcagg
     2881 gaacaaagag ttgtgtggca gagtggttgg atcagattgc aagatcgagg gaactaagtt
     2941 gaggctggag gtgctgttcc agggaccagg gagcgacaaa actcacacat gcccaccgtg
     3001 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     3061 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     3121 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     3181 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     3241 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     3301 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     3361 caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt
     3421 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     3481 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     3541 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     3601 tgagggtctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatctag
     3661 agggcccttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     3721 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3781 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3841 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3901 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3961 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     4021 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     4081 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4141 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6301 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6361 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6421 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6481 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6541 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6601 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6661 ggtaacgcca gggttttccc agtcacgacg tt
//
