LOCUS       sequence-274565-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G49770 X027_pECIA14 should be used with bait vector 
            AT5G49770 X027_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  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: AT5G49770 X027_pECIA14
FEATURES             Location/Qualifiers
     source          1..6824
                     /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             2120..2143
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3818..3841
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3857..3874
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3897..3914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3947..4081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3971..3990)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4025..4044
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4225..4241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4225..4241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4238..4260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4249..4265
                     /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(4273..4279,4280..4297,4298..4303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4318..4339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4456..4473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4627..5215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4707..4726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5386..6177,6178..6246))
                     /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     6009..6028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6247..6351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6419..6437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6475..6497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6597..6616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6810..6824,1..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 acagatggtt cagattttac
      541 tgctttacaa gccctgaaga acgaatggga cacgttatcc aagagttgga aaagctctga
      601 tccttgtgga accgaatggg ttggaattac atgtaacaac gataaccgtg ttgtttcaat
      661 atcattaacc aaccgtaact tgaaaggaaa gcttcctacc gaaatttcga cattgtctga
      721 attgcaaacc ttggatttga ctggcaatcc tgaattgtct ggaccgcttc cagcaaatat
      781 cggtaacctc aggaagttga ctttcttgag ccttatggga tgtgctttta atggtccaat
      841 ccctgattcc atagggaatc ttgaacaact tacaagactc tctctgaatt taaataaatt
      901 tagtggaaca attccggctt ccatgggacg gttatcgaaa ctatattggt tcgacatagc
      961 tgacaatcag ttagaaggaa agcttccagt ttctgatggg gcttctttac ctggacttga
     1021 catgcttctt caaactgggc attttcattt tggtaataac aagctttcag gcgaaatccc
     1081 agagaagctc tttagctcag aaatgacttt gctacatgtg ctgtttgatg gaaaccaatt
     1141 caccggtagc atcccagaga gcctcggcct agttcaaaat ttgactgtgt tacgtcttga
     1201 taggaataga ctaagtggag acattccttc aagtcttaat aatctcacaa atcttcaaga
     1261 actgcatttg tccgacaaca aatttacagg tagtcttcca aatttgacca gcttgaccag
     1321 tctctacaca ttggatgtga gcaataaccc gttggcgtta tcaccggttc catcatggat
     1381 ccctttctta aactctttgt caacattaag gttggaagat atccaacttg atggtccagt
     1441 accaacctcc ctttttagcc ctcttcaatt gcaaactgtt tccctaaagc acaatcttat
     1501 aaacacaaca ttggaccttg gtaccaacta tagcaaacaa ttggattttg tggatttacg
     1561 agacaacttc ataactggtt ataaatcacc agctaataac cccgtcaatg taatgttggc
     1621 agataatcaa gtgtgccaag acccggcgaa tcagctgagt ggctactgca atgcagtcca
     1681 accaaattcc acattttcta ccctaacaaa atgtggtaat cattgtggta aaggtaagga
     1741 accgaatcaa ggctgccact gtgtgtatcc acttacagga gtattcacct taaggtctcc
     1801 ttcattctcc ggattttcaa acaacagtaa cttcctaaag tttggggaga gtttaatgac
     1861 tttctttaag aatggcaagt atcctgtaga ctctgtggcc atgagaaaca taagcgagaa
     1921 tccaactgat tatcatcttt tgataaatct cttgatcttt ccatcgggac gcgaccgctt
     1981 taatcagacg gaaatggact ctattaattc tgcatttacc attcaggatt ataagcctcc
     2041 tccaagattt ggtccttata ttttcgtcgc cgatcagtac aaaacattct ctgatttaga
     2101 agattccaag acagtaagtc tggaggtgct gttccaggga ccaagcggca atcccggcgg
     2161 cgatctggcc ccccagatgc tgcgcgagct gcaggagacc aatgccgccc tgcaggacgt
     2221 gcgcgagctg ctgcgccacc gcgtgaagga gatcaccttc ctgaagaaca ccgtgatgga
     2281 gtgcgacgcc tgcggcatgc agccctctag aggctccggc tccatcatcc cagttgagga
     2341 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     2401 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     2461 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     2521 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     2581 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     2641 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     2701 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     2761 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     2821 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     2881 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtggag gccgaaagta
     2941 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     3001 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     3061 tgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     3121 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     3181 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     3241 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     3301 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     3361 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     3421 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     3481 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     3541 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     3601 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     3661 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     3721 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     3781 gcacccgggt tctagaggaa gcacctccgg cgataacgat tacaaggatg atgatgataa
     3841 ggccagcggc accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3901 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3961 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4021 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4081 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4141 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     4201 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4261 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4321 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4381 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4441 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4501 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4561 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5281 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5341 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6061 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6361 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6421 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6481 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6541 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6601 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6661 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6721 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6781 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
