LOCUS       sequence-275971-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector HSL2 D04_pECIA14 should be used with bait vector HSL2 
            D04_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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: HSL2 D04_pECIA14
FEATURES             Location/Qualifiers
     source          1..7040
                     /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             2336..2359
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4034..4057
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4073..4090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4113..4130)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4163..4297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4241..4260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4441..4457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4441..4457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4454..4476)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4465..4481
                     /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(4489..4495,4496..4513,4514..4519))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4534..4555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4672..4689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4843..5431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4923..4942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5602..6393,6394..6462))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNIGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6225..6244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6463..6567)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6635..6653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6691..6713)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6813..6832
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7026..7040,1..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 tccaatggag acgctgagat
      541 attgagtaga gttaaaaaga cccgactttt cgaccccgat ggaaatttac aagattgggt
      601 cataaccgga gataatcgga gtccatgtaa ttggacggga atcacatgcc acatcagaaa
      661 aggtagctcc ctcgccgtca ctaccattga tctctccggc tataatatct ccggtggctt
      721 tccctacgga ttctgtcgta tccgtacact catcaacatc actctttctc aaaacaatct
      781 caatggtacg attgattctg ctcctctctc cctctgttct aaacttcaga atttgattct
      841 caatcaaaac aacttctccg gtaaattacc ggaattctca ccggagtttc gtaaattacg
      901 agtcctcgaa ttggaatcaa acctcttcac cggtgagatt cctcaaagtt acgggagact
      961 cactgctctg caagttctga atcttaatgg taacccgctc agtggaatcg ttccggcgtt
     1021 tttgggttat ctgactgagt taactcgtct tgatctcgct tacatcagtt ttgatcctag
     1081 tccgattcca tcaaccttgg ggaacttgtc gaatctgact gatcttcggc taactcactc
     1141 gaacctcgtc ggagaaattc ctgattcgat catgaatctg gtgttgttag agaatcttga
     1201 tttagctatg aatagtctca ccggagaaat acctgagagt atcggaagac tcgaatcggt
     1261 ttaccagatt gagctctacg ataaccggtt atctggaaaa ttaccggaga gtatcggaaa
     1321 tttaaccgaa ttgaggaatt ttgatgtctc gcagaataat ctaaccggtg aactaccgga
     1381 aaagatcgct gctctgcaac ttatctcttt caatctcaat gataatttct tcaccggagg
     1441 attaccagat gtcgtagctt tgaatcctaa tctcgttgaa ttcaaaatct tcaacaacag
     1501 tttcacgggg acgttaccaa ggaatctcgg gaaattctca gaaatctctg aattcgatgt
     1561 ctcgacgaac agattctccg gtgaattgcc gccgtatttg tgctacagaa gaaaacttca
     1621 gaagattatc accttcagca atcaattaag cggcgaaatt ccggaatctt acggcgattg
     1681 tcattcgctt aattacattc gtatggcgga taacaaactc tccggcgaag ttccggctag
     1741 gttttgggaa cttcctctta ctcgtcttga gctagccaac aacaatcaat tacaaggttc
     1801 gattcctcct tcgatttcca aagctcgtca tctatctcag cttgaaatct ccgctaacaa
     1861 cttctccggt gtgattcccg tcaaactttg tgatctccgt gatctcagag tcatcgatct
     1921 tagccgcaac agtttcttag gatcaattcc gtcttgcatc aacaaattga agaatctaga
     1981 gagagtagag atgcaggaga acatgctcga cggcgagatt ccgagttcag tgagttcgtg
     2041 caccgagtta accgaattaa atctctccaa caaccgttta cgaggcggga taccaccgga
     2101 actcggtgat ttaccggttt taaactacct ggatctctct aacaaccaac tcaccggtga
     2161 gattccggcg gagctgttga ggctcaagct taatcaattc aacgtctccg ataacaaact
     2221 ctatggtaag attccttctg gatttcagca agatattttt cgacccagtt tcttaggtaa
     2281 cccgaatctc tgtgccccaa atttggatcc gattagacct tgccgatcca aacggctgga
     2341 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     2401 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     2461 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     2521 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     2581 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     2641 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     2701 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     2761 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     2821 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     2881 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     2941 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     3001 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     3061 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     3121 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     3181 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     3241 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     3301 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     3361 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     3421 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     3481 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     3541 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     3601 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     3661 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     3721 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     3781 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     3841 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     3901 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     3961 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     4021 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     4081 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     4141 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     4201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4321 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     4381 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     4441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5581 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5641 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5701 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5761 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5821 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5881 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5941 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6001 ccctatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6061 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6121 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6181 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6241 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6301 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6361 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6421 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6481 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6541 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6601 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6661 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6721 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6781 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6841 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6901 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6961 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     7021 gttttcccag tcacgacgtt
//
