LOCUS       sequence-298833-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector GSO2/EDA23 X079_pECIA2 should be used with prey vector 
            GSO2/EDA23 X079_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  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 6806)
  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: GSO2/EDA23 X079_pECIA2
FEATURES             Location/Qualifiers
     source          1..6806
                     /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             3059..3082
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3788..3829
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3839..3856
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3879..3896)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3929..4063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3953..3972)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4007..4026
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4207..4223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4207..4223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4220..4242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4231..4247
                     /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(4255..4261,4262..4279,4280..4285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4300..4321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4438..4455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4609..5197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4689..4708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5368..6159,6160..6228))
                     /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     5991..6010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6229..6333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6401..6419
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6457..6479)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6579..6598
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6792..6806,1..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 caaagagacg atcttcaaac
      541 tcttctcgaa ctgaagaact cttttatcac aaacccaaaa gaagaagacg ttctccgaga
      601 ttggaattcc ggtagtccca gttactgcaa ctggaccggc gtcacatgcg gtggtcgtga
      661 aatcatcggt ttaaatctct ccggtttagg tttaaccggt tcaatttctc cttcgatcgg
      721 ccggtttaac aacctaatcc acatcgattt gtcttcgaac cgtcttgtcg gtcccatccc
      781 aacgactctc tctaacctct cctcttcatt ggaatcgttg catctcttct ctaacctact
      841 cagtggcgat ataccgagtc aactcggctc acttgtgaat ctcaaatcgt taaaactcgg
      901 agacaacgaa ctcaacggaa caatcccgga gacgtttggg aacctcgtca acctccagat
      961 gcttgctttg gcttcgtgta gactcaccgg tcttataccg agtcgattcg gtagactcgt
     1021 tcagctccag actttgatac tacaagacaa cgaactcgaa ggaccgattc cggcggaaat
     1081 cggaaactgc accagcctcg ctttgtttgc cgcggcgttt aatcgtctca acggttcgtt
     1141 accggcggag ctgaatcgac ttaagaacct ccagacgctg aatttgggag ataacagttt
     1201 ctccggtgag ataccgagtc aactcggtga tttggttagt atccaatacc ttaatttgat
     1261 tggtaatcag ttacagggtt tgattccaaa gagattaaca gaattagcga atcttcaaac
     1321 acttgatttg tcttctaata atctcaccgg agtgatacac gaagagttct ggagaatgaa
     1381 tcagcttgag ttcttggtat tagcgaaaaa tcgtctctct ggttcgttac caaagactat
     1441 atgttctaac aacacaagct tgaagcaact gtttctgtct gaaactcagc tttccggcga
     1501 aattccagcg gaaatcagca actgtcagtc attaaaattg cttgatttgt cgaacaatac
     1561 gcttacaggt caaatcccgg attcgttgtt tcagctcgtc gaactcacga atctttatct
     1621 taacaacaat agcttggagg gtacgttatc ttcatcaata tcaaacctca cgaatctaca
     1681 agagtttact ctctatcaca ataacttgga aggcaaggta cctaaagaaa tcggatttct
     1741 tggtaaattg gagattatgt atctgtatga gaatcggttt tcgggtgaaa tgccggttga
     1801 gattgggaat tgcacgagac tgcaggagat tgattggtac gggaatcgtt taagtggaga
     1861 gattccttct tcaattggaa gattgaaaga tcttactcgt cttcacttga gagagaacga
     1921 gcttgtaggt aacattcccg ccagtttagg taactgtcat caaatgacgg ttatcgattt
     1981 agcggataac cagctctcgg gttcaattcc ttcctcgttc gggttcttaa cggcattaga
     2041 actgtttatg atctacaaca attctcttca aggaaatctt ccggattcac ttatcaacct
     2101 caagaacctc acaagaatca atttttcaag caacaagttc aatggttcga taagtccatt
     2161 gtgcggttca agctcgtatc tctcgtttga tgtcacggaa aacggatttg agggagatat
     2221 acctcttgag ctaggtaagt ctacgaatct tgatcggtta aggctgggga agaatcagtt
     2281 cacaggaaga atccctcgga catttgggaa gatcagtgag ctttctttat tggatatttc
     2341 aagaaactct ctctcaggaa tcataccggt ggagctcggt ttgtgcaaga agctgacaca
     2401 cattgatctc aacaataact atctttcagg ggtgatacct acatggcttg gaaagcttcc
     2461 attgttggga gagctaaagc tttcttctaa taagtttgtt gggtctctcc ctactgagat
     2521 tttcagctta accaatattc ttacgctatt tcttgatggt aactcactca acggctcgat
     2581 cccgcaagag attggtaact tgcaagctct caatgcgttg aacctcgaag agaatcagct
     2641 ttcaggtcct ctgccttcta caatagggaa actgagcaag cttttcgagc ttcggttatc
     2701 gagaaacgct ttaaccggag agattcctgt tgagattgga cagctgcaag atcttcaaag
     2761 tgctttagat ctcagctaca acaactttac cggacgtata ccgtctacga tttcgacgtt
     2821 acctaagctt gaatctcttg atctgtctca caatcagctt gtaggagaag ttcctggcca
     2881 aatcggtgac atgaaaagct tgggatatct taacctctct tacaacaacc ttgagggaaa
     2941 gttaaagaaa cagttctcta gatggcaagc tgatgcgttt gtaggcaatg caggtctttg
     3001 tggaagccct cttagtcatt gcaacagagc tggatcaaag aatcaacgaa gtcttagtct
     3061 ggaggtgctg ttccagggac cagggagcga caaaactcac acatgcccac cgtgcccagc
     3121 acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct
     3181 catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc
     3241 tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc
     3301 gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca
     3361 ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc
     3421 catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg tgtacaccct
     3481 gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg
     3541 cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta
     3601 caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac
     3661 cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggg
     3721 tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaat ctagagggcc
     3781 cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     3841 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3901 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3961 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4021 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4081 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4141 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     4201 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6361 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6421 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6481 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6541 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6601 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6661 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6721 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6781 gccagggttt tcccagtcac gacgtt
//
