LOCUS       sequence-298762-        5618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BARK1 X073_pECIA2 should be used with prey vector BARK1 
            X073_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5618)
  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 5618)
  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: BARK1 X073_pECIA2
FEATURES             Location/Qualifiers
     source          1..5618
                     /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             1871..1894
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2600..2641
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2651..2668
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2691..2708)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2741..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2765..2784)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2819..2838
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3019..3035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3019..3035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3032..3054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3043..3059
                     /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(3067..3073,3074..3091,3092..3097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3112..3133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3250..3267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3421..4009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3501..3520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4180..4971,4972..5040))
                     /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     4803..4822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5041..5145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5213..5231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5269..5291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5391..5410
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5604..5618,1..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 tccgtcgctg atgaccaaac
      541 agcaatgctc gctttagcta aatcattcaa tcctccgccg tcagattggt cttccaccac
      601 cgatttctgc aaatggtccg gcgtacgatg taccggcggt cgtgtcacaa ccatcagcct
      661 cgctgataaa tctctcaccg gattcatcgc acctgagatc tcgactcttt cagagctcaa
      721 atctgtttct attcaacgta acaaactctc cggtacaatc ccttcgtttg ctaaactctc
      781 gtctcttcaa gagatctata tggatgagaa taatttcgtc ggagttgaaa ccggagcttt
      841 cgccggactt actagtctcc agatcttgag tttaagcgat aacaacaaca ttactacttg
      901 gagtttccca tcggagctcg tcgattcgac ttctctcact acgatttatc tcgataacac
      961 aaacatcgcc ggagttttgc ctgacatttt cgattcctta gcttctcttc aaaatctccg
     1021 gttatcttac aacaacatca ccggcgtttt accaccgtcg cttggaaaat cttcgattca
     1081 gaatctttgg atcaacaatc aagacctagg aatgtcagga acgatcgaag ttctttcgag
     1141 tatgacgtcc ttgtcacaag cttggcttca caagaatcac ttctttggac cgattccaga
     1201 tctctccaag agcgagaatc tcttcgatct acagctccgg gataatgatt taaccggaat
     1261 agtacctccg acgcttctta cactcgctag cttgaagaat atttctctag ataacaataa
     1321 atttcaagga cctcttcctt tgttttcacc ggaggttaaa gtaacaatag accacaacgt
     1381 tttttgtact actaaagctg gacaaagctg tagtcctcag gtgatgacgc ttttagcggt
     1441 ggctggaggt ttgggatatc cttcaatgtt ggctgagtct tggcaaggtg atgatgcttg
     1501 tagtggttgg gcttatgtta gttgtgattc agctgggaag aatgttgtca cgttgaatct
     1561 tgggaaacat ggattcaccg ggtttatatc tccggcgatt gcgaatctta cttcattgaa
     1621 gagtctttat cttaatggta acgatttgac tggtgttatt cctaaggagt tgacatttat
     1681 gactagtttg cagttaattg atgtctcgaa taacaatctt agaggggaga tacctaagtt
     1741 tccggctacg gtgaagttta gttacaaacc ggggaatgct ttgttgggaa ctaatggtgg
     1801 agatggttcg agtcctggaa ctggtggtgc tagtggtggt cctggtgggt cttctggtgg
     1861 tggtggtagt ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     1921 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     1981 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     2041 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     2101 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     2161 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     2221 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     2281 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     2341 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     2401 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2461 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     2521 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     2581 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2641 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2701 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     2761 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2821 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2881 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2941 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     3001 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3061 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3121 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3181 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3241 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3301 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3841 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4081 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4261 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4381 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4621 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4681 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4861 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5041 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5101 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5161 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5221 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5281 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5341 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5401 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5461 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5521 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5581 aagttgggta acgccagggt tttcccagtc acgacgtt
//
