LOCUS       sequence-298783-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector HSL2 D04_pECIA2 should be used with prey vector HSL2 
            D04_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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 6083)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6083
                     /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             3065..3106
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3116..3133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3156..3173)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3206..3340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3230..3249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3284..3303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3484..3500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3484..3500)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3497..3519)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3508..3524
                     /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(3532..3538,3539..3556,3557..3562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3577..3598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3715..3732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3886..4474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3966..3985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4645..5436,5437..5505))
                     /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     5268..5287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5506..5610)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5678..5696
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5734..5756)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5856..5875
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6069..6083,1..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 cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     2401 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     2461 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     2521 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     2581 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     2641 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     2701 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     2761 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     2821 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     2881 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     2941 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     3001 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     3061 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     3121 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     3181 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     3241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3301 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3361 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3421 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     3481 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3541 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3601 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3661 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3721 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3781 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3841 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3901 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3961 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4021 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4081 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4141 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4201 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4261 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4321 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4381 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4441 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4501 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4561 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4621 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4741 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4801 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5161 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5521 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5581 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5641 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5701 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5761 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5821 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5881 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     5941 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6001 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6061 agggttttcc cagtcacgac gtt
//
