LOCUS       sequence-298816-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector IKU2 X076_pECIA2 should be used with prey vector IKU2 
            X076_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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: IKU2 X076_pECIA2
FEATURES             Location/Qualifiers
     source          1..6044
                     /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             2297..2320
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3026..3067
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3077..3094
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3117..3134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3167..3301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3191..3210)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3245..3264
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3445..3461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3445..3461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3458..3480)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3469..3485
                     /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(3493..3499,3500..3517,3518..3523))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3538..3559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3676..3693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3847..4435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3927..3946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4606..5397,5398..5466))
                     /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     5229..5248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5467..5571)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5639..5657
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5695..5717)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5817..5836
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6030..6044,1..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 tccaaccaca gtgaagaagt
      541 cgagaatctt ctcaaactga aatccacctt tggagaaaca aaatccgacg acgtctttaa
      601 gacctggact caccggaatt cggcatgtga attcgccgga atcgtatgta attccgacgg
      661 aaacgtcgtc gagatcaacc ttggaagtcg gagtttgatc aaccgagacg acgacggtag
      721 gttcaccgat cttcctttcg attcaatctg cgatttgaag ttgctggaga agcttgttct
      781 aggaaacaac tctttacgtg gtcagattgg tacgaatctc gggaaatgta atcgcttgag
      841 atacttagac ctcgggatca acaatttctc cggtgagttt ccggcgattg attcgttaca
      901 attactcgag tttttgagct taaacgcttc tggaatctct ggaatattcc catggagttc
      961 tctaaaagac ctgaaacgat tgagtttcct aagcgttgga gataaccgat tcggttcgca
     1021 tccatttcct agagagattc taaatctcac tgccttgcaa tgggtttatc tgagtaacag
     1081 cagcatcact ggtaagattc cagagggaat caaaaacctt gttcgtctac aaaacttgga
     1141 gctctctgat aatcagatct ctggtgaaat tccaaaggag attgttcagc tcaaaaacct
     1201 aaggcagctt gagatttata gtaatgactt gaccggtaag ttaccattgg gttttaggaa
     1261 tttgacgaat ctgaggaatt tcgacgcctc taacaactcc ttggaaggtg atttatcaga
     1321 gctgcggttc ttaaagaatc ttgtgtctct tggaatgttc gaaaaccgat taaccggtga
     1381 gattccaaaa gagtttggag atttcaagag tttagctgct ttgtctcttt acagaaacca
     1441 gctcacaggg aagcttccga ggagactcgg ttcttggaca gcttttaagt acatcgacgt
     1501 gtctgagaac ttcttggaag gtcagattcc tccttatatg tgcaagaaag gcgtaatgac
     1561 gcatcttctc atgttgcaga acagattcac aggacaattt ccagagagtt atgctaaatg
     1621 taagactctg attcgtcttc gtgtaagtaa caactctctc tctggaatga ttccttcagg
     1681 gatttggggt cttcctaatc tacagtttct tgaccttgct tcaaactatt ttgaaggaaa
     1741 cctcactggt gatattggta atgccaagtc tctaggttct ttagatttga gcaataatag
     1801 attttcaggt tctctaccgt ttcagatctc aggagctaac tcgttagtgt cggttaatct
     1861 tcgaatgaat aagttttccg ggatagttcc tgaatctttt ggtaagctta aggagctttc
     1921 aagtctcatt cttgaccaga ataatctatc tggtgctata ccaaagtcac taggcttatg
     1981 cacatctttg gttgatctaa actttgctgg aaactcactc tctgaggaaa tcccggaaag
     2041 tttaggatct ttaaagttat tgaattccct gaatctgtca ggaaacaaat tatcaggaat
     2101 gattcctgtt ggtttgtcgg ctctgaagct aagcttactt gacttatcca acaaccaatt
     2161 gaccggttct gttcctgaat ctctcgtaag tggaagcttt gaaggaaatt caggactatg
     2221 cagctcaaag atcagatatc ttcgtccatg tccgcttgga aaacctcaca gccaagggaa
     2281 gaggaaacat ttgtcactgg aggtgctgtt ccagggacca gggagcgaca aaactcacac
     2341 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     2401 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     2461 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     2521 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     2581 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     2641 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     2701 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     2761 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     2821 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     2881 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     2941 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     3001 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3061 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3121 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3181 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3241 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3301 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3361 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     3421 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5581 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5641 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5701 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5761 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5821 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5881 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     5941 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6001 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
