LOCUS       sequence-296819-        5012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G43020 K6_pECIA2 should be used with prey vector 
            AT5G43020 K6_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5012)
  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 5012)
  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: AT5G43020 K6_pECIA2
FEATURES             Location/Qualifiers
     source          1..5012
                     /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             1265..1288
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1994..2035
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2045..2062
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2085..2102)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2135..2269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2159..2178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2213..2232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2413..2429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2413..2429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2426..2448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2437..2453
                     /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(2461..2467,2468..2485,2486..2491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2506..2527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2644..2661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3574..4365,4366..4434))
                     /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     4197..4216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4435..4539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4607..4625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4663..4685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4785..4804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4998..5012,1..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 caaagcgcgt
      541 gtttcattct caccgtgacg tttctgcttt gttacgtttt aaatccaaag ctgatttatg
      601 gaacaagata aacacgagct ctcatttttg tcaatggtgg ggtgttacgt gttatggaaa
      661 cagagttgtt cgtttggtta ttgaagatct ttatcttgga gggcgactca ttcctgactc
      721 agttaataaa ctcgaccagc ttcgagttct gagtctcaag aacacttcct taaccggacc
      781 gttaccagat ttctccggtt tggttaatct caaatcatta ttcctcgatc acaactcttt
      841 ttccggttcg tttcctctct cggttctcgc ttttcaccgg ttaagaactc tcgatttctc
      901 cttcaataac ctcaccggtc caatcccatc cggtttagtt ctctccgacc ggttgattta
      961 tctccgactt gattcgaatc ggttcaacgg tcccgtcccg cctttaaacc aatccacgct
     1021 tcatacattc aatgtttcgg ttaacaactt aaccggtgct gtaccggtga caaccgttct
     1081 gttacgattc ggtatttctt catttctcaa aaaccctaat ctctgcggtg agattgttca
     1141 caaagaatgc aatccgagag ccaaattctt cactccggta acggcggcgc cgtcaccgaa
     1201 gatggttctc ggtcaaatag cgcaaatagg cggcgcaaga ttatcccggc caagtcaaaa
     1261 caaactggag gtgctgttcc agggaccagg gagcgacaaa actcacacat gcccaccgtg
     1321 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     1381 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     1441 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     1501 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     1561 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     1621 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     1681 caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt
     1741 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     1801 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     1861 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     1921 tgagggtctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatctag
     1981 agggcccttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     2041 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2101 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     2161 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2221 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2281 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2341 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     2401 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2461 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2521 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3541 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3601 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3661 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3721 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3781 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3841 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3901 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3961 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4021 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4081 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4141 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4201 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4261 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4321 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4381 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4561 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4621 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4681 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4741 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4801 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4861 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4921 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4981 ggtaacgcca gggttttccc agtcacgacg tt
//
