LOCUS       sequence-298829-        6068 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector HAESA D03_pECIA2 should be used with prey vector HAESA 
            D03_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6068)
  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 6068)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6068
                     /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             3050..3091
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3101..3118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3141..3158)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3191..3325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3215..3234)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3269..3288
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3469..3485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3469..3485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3482..3504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3493..3509
                     /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(3517..3523,3524..3541,3542..3547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3562..3583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3700..3717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3871..4459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3951..3970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4630..5421,5422..5490))
                     /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     5253..5272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5491..5595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5663..5681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5719..5741)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5841..5860
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6054..6068,1..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 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     2401 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     2461 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     2521 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     2581 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     2641 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     2701 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     2761 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     2821 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     2881 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     2941 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     3001 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     3061 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     3121 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3181 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3241 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3301 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3361 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3421 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3481 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5701 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5761 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5821 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5881 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5941 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6001 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6061 acgacgtt
//
