LOCUS       sequence-296820-        4976 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector AT5G58300 K8_pECIA2 should be used with prey vector 
            AT5G58300 K8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  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: AT5G58300 K8_pECIA2
FEATURES             Location/Qualifiers
     source          1..4976
                     /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             1229..1252
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1958..1999
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2009..2026
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2049..2066)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2099..2233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2123..2142)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2177..2196
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2377..2393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2377..2393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2390..2412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2401..2417
                     /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(2425..2431,2432..2449,2450..2455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2470..2491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2608..2625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2779..3367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2859..2878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3538..4329,4330..4398))
                     /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     4161..4180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4399..4503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4571..4589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4627..4649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4749..4768
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4962..4976,1..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 ctgaactcgg ataggcaagc
      541 tcttcttgca tttgctgctt ctgttcctca tctcagacga ctcaactgga attccactaa
      601 ccatatctgc aaatcatggg ttggcgtcac ctgcacttct gatggtacta gtgttcacgc
      661 tcttcgcctt ccaggaattg ggctcttggg accaatccct ccaaacacac ttggtaaact
      721 agaatcactc aggattctca gtttgagatc aaacctcctc agtggtaacc ttcctcctga
      781 cattcactct ctcccttcac tagattacat ctatctccaa cataacaact tctctggtga
      841 ggttccttcc tttgtgtccc gacagctcaa tatcttagat ctctcattta actctttcac
      901 cgggaagata ccagcaacgt tccaaaatct gaagcaattg acagggttga gcctccaaaa
      961 taacaaactt tctggaccag tacctaatct tgatacagta agtctgagac gtttaaactt
     1021 gagcaacaat catcttaatg gttcaatacc ctctgcactt ggtggctttc ccagttcatc
     1081 tttttccgga aacacattgt tatgtggact tcctctgcaa ccctgtgcca cctcctctcc
     1141 ccctccttcg ctaaccccac acatttccac gcctccattg ccgccatttc ctcacaaaga
     1201 aggttcaaaa aggaaactac atgtaagcct ggaggtgctg ttccagggac cagggagcga
     1261 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     1321 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     1381 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     1441 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     1501 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     1561 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     1621 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     1681 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     1741 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     1801 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     1861 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     1921 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     1981 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2041 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2101 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2161 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2221 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2281 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     2341 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2401 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2461 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2521 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4501 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4561 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4621 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4681 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4741 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4801 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     4861 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     4921 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
