LOCUS       sequence-296763-        4862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G10290 I2_pECIA2 should be used with prey vector 
            AT5G10290 I2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4862)
  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 4862)
  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: AT5G10290 I2_pECIA2
FEATURES             Location/Qualifiers
     source          1..4862
                     /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             1115..1138
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1844..1885
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1895..1912
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1935..1952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1985..2119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2009..2028)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2063..2082
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2263..2279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2263..2279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2276..2298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2287..2303
                     /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(2311..2317,2318..2335,2336..2341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2356..2377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2494..2511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2665..3253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2745..2764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3424..4215,4216..4284))
                     /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     4047..4066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4285..4389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4457..4475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4513..4535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4635..4654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4848..4862,1..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 tctccagatg ctcaagggga
      541 tgcactgttt gcgttgagga tctccttacg tgcattaccg aatcagctaa gtgactggaa
      601 tcagaaccaa gttaatcctt gcacttggtc ccaagttatt tgtgatgaca aaaactttgt
      661 cacttctctt acattgtcag atatgaactt ctcgggaacc ttgtcttcaa gagtaggaat
      721 cctagaaaat ctcaagactc ttactttaaa gggaaatgga attacgggtg aaataccaga
      781 agactttgga aatctgacta gcttgactag tttggatttg gaggacaatc agctaactgg
      841 tcgtatacca tccactatcg gtaatctcaa gaaacttcag ttcttgacct tgagtaggaa
      901 caaacttaat gggactattc cggagtcact cactggtctt ccaaacctgt taaacctgct
      961 gcttgattcc aatagtctca gtggtcagat tcctcaaagt ctgtttgaga tcccaaaata
     1021 taatttcacg tcaaacaact tgaattgtgg cggtcgtcaa cctcaccctt gtgtatccgc
     1081 ggttgcccat tcaggtgatt caagcaagcc taaactggag gtgctgttcc agggaccagg
     1141 gagcgacaaa actcacacat gcccaccgtg cccagcacct gaactcctgg ggggaccgtc
     1201 agtcttcctc ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt
     1261 cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt
     1321 ggacggcgtg gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac
     1381 gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta
     1441 caagtgcaag gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc
     1501 caaagggcag ccccgagaac cacaggtgta caccctgccc ccatcccggg aggagatgac
     1561 caagaaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt
     1621 ggagtgggag agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga
     1681 ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca
     1741 ggggaacgtc ttctcatgct ccgtgatgca tgagggtctg cacaaccact acacgcagaa
     1801 gagcctctcc ctgtctccgg gtaaatctag agggcccttc gaaggtaagc ctatccctaa
     1861 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
     1921 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     1981 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2041 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2101 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2161 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     2221 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     2281 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2341 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2401 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2461 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2521 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2581 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3301 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3361 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3421 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3481 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3601 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3841 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4081 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4261 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4381 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4441 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4501 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4561 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4621 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     4681 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     4741 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4801 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4861 tt
//
