LOCUS       sequence-296773-        4940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G60630 I8_pECIA2 should be used with prey vector 
            AT1G60630 I8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4940)
  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 4940)
  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: AT1G60630 I8_pECIA2
FEATURES             Location/Qualifiers
     source          1..4940
                     /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             1193..1216
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1922..1963
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1973..1990
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2013..2030)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2063..2197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2087..2106)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2141..2160
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2341..2357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2341..2357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2354..2376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2365..2381
                     /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(2389..2395,2396..2413,2414..2419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2434..2455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2823..2842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3502..4293,4294..4362))
                     /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     4125..4144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4363..4467)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4535..4553
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4591..4613)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4713..4732
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4926..4940,1..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 agcgatgttg aagctctact
      541 gagtttgaaa tcatccattg atccttcaaa ttcgattcca tggcgaggaa cagatccttg
      601 caactgggaa ggagtcaaga aatgcatgaa aggaagagtc tcaaagcttg ttcttgagaa
      661 tttgaacctc agtggctcac tcaacggcaa gagtttgaac caattggatc aacttagagt
      721 tcttagtttc aaaggaaact ctctctctgg ctcaatccct aacctatctg gtttagtcaa
      781 cctcaaatct ctttatctca acgataataa cttctccggt gagttccctg aatcactgac
      841 gtctctacac cgattgaaaa cggttgttct ttccaggaat cggttttccg gcaagattcc
      901 tagctcgttg ctccgactct cccgccttta cacgttttac gttcaagaca atctcttctc
      961 tggttccatt cctcctctca accaagcaac tctcagattc tttaatgtct cgaataatca
     1021 actctctggt catatcccac caacacaagc tctgaatcgg ttcaatgagt cgtcgtttac
     1081 tgacaacatt gctctctgcg gtgatcagat tcaaaactca tgcaatgaca ccactggaat
     1141 cacttcaact ccatcagcaa aaccagctat ccctgtagcc aaaacacgta gcctggaggt
     1201 gctgttccag ggaccaggga gcgacaaaac tcacacatgc ccaccgtgcc cagcacctga
     1261 actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat
     1321 ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt
     1381 caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga
     1441 ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg
     1501 gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga
     1561 gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc
     1621 atcccgggag gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta
     1681 tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac
     1741 cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga
     1801 caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg agggtctgca
     1861 caaccactac acgcagaaga gcctctccct gtctccgggt aaatctagag ggcccttcga
     1921 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     1981 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     2041 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     2101 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2161 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2221 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2281 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     2341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3181 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4501 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4561 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4621 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4681 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4741 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     4801 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     4861 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     4921 gttttcccag tcacgacgtt
//
