LOCUS       sequence-296423-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector HAESA D03_pECIA14 should be used with bait vector HAESA 
            D03_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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: HAESA D03_pECIA14
FEATURES             Location/Qualifiers
     source          1..7025
                     /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             2321..2344
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4019..4042
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4058..4075
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4098..4115)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4148..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4172..4191)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4226..4245
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4426..4442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4426..4442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4439..4461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4450..4466
                     /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(4474..4480,4481..4498,4499..4504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4519..4540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4657..4674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4828..5416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4908..4927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5587..6378,6379..6447))
                     /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     6210..6229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6448..6552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6620..6638
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6676..6698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6798..6817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7011..7025,1..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 tctctaaacc aagatgccac
      541 cattctccgg caagccaaac tgggcttatc cgacccggcc cagtccctat cttcatggtc
      601 cgacaacaac gacgtcaccc catgcaagtg gctaggcgtg agctgcgacg ccacgtcaaa
      661 cgttgtctcc gtcgatctct caagcttcat gctcgtcgga cctttcccct ccatcctctg
      721 ccaccttcct tctctccatt ccctctctct atataataac tccatcaatg gctctctctc
      781 cgccgacgac ttcgatacgt gtcacaatct catcagtctt gatttatcgg agaatctatt
      841 ggttggatcc atcccaaagt cacttccttt caatctccca aacctcaagt tccttgaaat
      901 ctccggaaac aacttatcag acacgatccc gtcgagcttc ggagagtttc gaaagcttga
      961 gtctttaaac ctcgccggta acttcctctc cggtacaatt cccgcctctc tcggtaacgt
     1021 cacgactctt aaagaactca aactcgctta caatttgttt tctccgagtc aaatcccgag
     1081 tcaactcgga aacctcacgg agctccaagt tctctggctc gccggctgca acctcgtcgg
     1141 tccgatacct ccgtctctgt cccggttaac tagtttggtt aacttggatt tgacatttaa
     1201 ccaactcaca ggatctatcc caagttggat cacgcagcta aaaaccgtcg agcaaatcga
     1261 actattcaac aactcgttct caggcgagtt accagaatcc atgggtaaca tgacgacgct
     1321 aaaaagattc gacgcgtcga tgaataagct gacaggaaaa atacccgaca acttgaatct
     1381 gttaaacctc gagtcactca atctcttcga gaacatgctc gaaggtcctt taccggagag
     1441 cataactcgc tccaaaacct tgtcagaact caagcttttc aacaacagac tcaccggagt
     1501 attaccgagt caactcggcg ctaactcacc gttacagtac gtggacttat cgtataaccg
     1561 attttccggc gaaataccgg cgaatgtctg cggtgaaggg aagctagagt atctaattct
     1621 tatagacaat tcgttttcag gagaaatctc taataatctc ggaaagtgta agagcttgac
     1681 ccgggtccga ttgagtaaca acaagctttc gggtcaaatc ccacatggat tctggggatt
     1741 gcctcgcttg tcacttctcg aactctctga caattcgttc accggaagta ttcctaagac
     1801 gatcatcggc gcgaagaatc tatcgaacct acgaatctca aagaatcgat tttcaggttc
     1861 aatccccaac gaaatcggat cgcttaacgg aataatcgag atttccggag cggagaatga
     1921 tttctccggt gagattcctg aaagcttggt gaaattgaag caattgagca ggcttgatct
     1981 cagtaagaat caactctccg gtgagattcc tagagaactt cgtggttgga agaatctaaa
     2041 cgagcttaat ttggctaata atcatctctc cggtgaaatc cctaaagaag tcggaatctt
     2101 acctgttctc aattaccttg acctttcaag caatcaattc tccggtgaga ttccgttgga
     2161 gttacagaat ctgaagctaa acgttttaaa cctttcgtat aatcacctct ccggtaaaat
     2221 ccctcctctc tacgctaata aaatctacgc acatgacttc atcggaaatc ccggtttatg
     2281 tgttgatctc gacggccttt gtcggaagat cacacggtct ctggaggtgc tgttccaggg
     2341 accaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     2401 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     2461 cctgaagaac accgtgatgg agtgcgacgc ctgcggcatg cagccctcta gaggctccgg
     2521 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     2581 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     2641 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     2701 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     2761 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     2821 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     2881 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     2941 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     3001 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     3061 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     3121 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     3181 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     3241 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     3301 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     3361 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     3421 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     3481 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     3541 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     3601 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     3661 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     3721 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     3781 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     3841 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     3901 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     3961 cgccggcacc accgacgccg cgcacccggg ttctagagga agcacctccg gcgataacga
     4021 ttacaaggat gatgatgata aggccagcgg caccggtcat catcaccatc accattgagt
     4081 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     4141 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4201 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4261 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4321 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     4381 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     4441 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4501 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5521 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6601 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6661 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6721 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6781 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6841 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6901 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6961 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7021 acgtt
//
