LOCUS       sequence-296427-        7763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector GSO2/EDA23 X079_pECIA14 should be used with bait vector 
            GSO2/EDA23 X079_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7763)
  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 7763)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..7763
                     /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             4757..4780
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4796..4813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4836..4853)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4886..5020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4910..4929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4964..4983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5164..5180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5164..5180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5177..5199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5188..5204
                     /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(5212..5218,5219..5236,5237..5242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5257..5278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6325..7116,7117..7185))
                     /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     6948..6967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7186..7290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7358..7376
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7414..7436)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7536..7555
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7749..7763,1..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 caagcggcaa tcccggcggc gatctggccc cccagatgct
     3121 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     3181 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
     3241 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     3301 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     3361 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     3421 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     3481 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     3541 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     3601 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     3661 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     3721 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     3781 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3841 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3901 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3961 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     4021 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     4081 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     4141 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     4201 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     4261 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     4321 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     4381 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     4441 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     4501 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     4561 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     4621 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     4681 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
     4741 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
     4801 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     4861 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     4921 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4981 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5041 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5101 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     5161 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5221 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5341 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7201 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7321 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7381 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7441 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7501 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7561 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7621 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7681 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7741 agggttttcc cagtcacgac gtt
//
