LOCUS       sequence-270147-        6653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G28990 N1_pECIA14 should be used with bait vector 
            AT2G28990 N1_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6653)
  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 6653)
  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: AT2G28990 N1_pECIA14
FEATURES             Location/Qualifiers
     source          1..6653
                     /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             1949..1972
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3647..3670
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3686..3703
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3726..3743)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3776..3910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3800..3819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3854..3873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4054..4070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4054..4070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4067..4089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4078..4094
                     /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(4102..4108,4109..4126,4127..4132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4147..4168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4285..4302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4456..5044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4536..4555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5215..6006,6007..6075))
                     /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     5838..5857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6076..6180)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6248..6266
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6304..6326)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6426..6445
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6639..6653,1..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 agtttagatt gtggattacc
      541 tagtgacgaa tctccttacg atgattcgtt taacggatta acattcacat cggattccac
      601 tttcatccaa acaggaaaaa ttgatagtgt cgataaagat cttaacataa acttatcaaa
      661 acaatatttg actcttagat acttcccaga aggaaaacgt aattgttata gtcttgatgt
      721 caagcgtggc acaacttatc tcatcgtggt tagctttgta tatggaaatt atgatggtct
      781 taaccgtgat ccaaacttcg atatccatct tggtcccaat aagtggaaaa gaatagattt
      841 ggatggagag aaagaaggta caagagaaga gatcatacac aaagccaggt caaactcttt
      901 ggatatttgt cttgttaaga caggagaaac tttgccaata atttcagcca tagaaatacg
      961 gccactaagg aataatacgt atgttacaca atctggttcg ctgatgatgt ctttccgggt
     1021 ttatcttagc aattcagatg catctataag gtatgctgat gacgtccatg atcgtatttg
     1081 gtctccattc aacggatctt cacatacaca tataacaact gatctcaata taaacaactc
     1141 aaatgcatat gagattccaa aaaatatact ccaaactgct gccataccaa gaaacgccag
     1201 tgcgccgctg attattacgt gggatccact acccattaat gcggaggtat acttgtacat
     1261 gcactttgcg gagattcaaa cccttgaagc caacgagacg aggcagtttg acgttatctt
     1321 gagaggaaac tttaatcatt caggatttag tccaaccaag ttaaaggtat ttacattata
     1381 cacagaagaa ccaatgaaat gtggttcaga aggttgttat ttacagctag taaaaacacc
     1441 gaactcaact ctgccacctc tgatcaacgc tattgaggca tacagtgtta tcgagttctc
     1501 acaattggaa acaagtctta gtgatgtgga cgctatcaag aacatcaaaa atacttataa
     1561 gctgaataaa atcacttggc aaggagatcc atgtctccct caagatttgt cgtgggaaag
     1621 tattagatgc acctacgtag atggctccac ttcaccaaca atcatttcct tagatttgtc
     1681 gaaaagtgga ttaaatggga gtatacccca aattttacaa aatttcacgc aattacaaga
     1741 actggactta tcaaataata gcttgactgg accggtgcct atcttccttg ccaacatgaa
     1801 aacgttatct ttaataaact taagtgggaa caatctcagt ggttcagttc ctcaagctct
     1861 tcttgacaaa gaaaaagaag gacttgtgtt aaaacttgaa ggaaatccag atttgtgcaa
     1921 atccagtttt tgcaataccg aaaagaaact ggaggtgctg ttccagggac caagcggcaa
     1981 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     2041 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     2101 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2161 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2221 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     2281 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     2341 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2401 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2461 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2521 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     2581 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     2641 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     2701 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     2761 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     2821 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     2881 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     2941 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     3001 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3061 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3121 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3181 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3241 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     3301 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3361 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3421 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3481 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3541 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     3601 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     3661 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3721 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3901 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3961 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     4021 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4081 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4141 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4201 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4261 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4321 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4861 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5161 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5221 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5281 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5341 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5401 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5461 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5521 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5581 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5641 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5701 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5761 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5821 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5881 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5941 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6001 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6061 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6121 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6181 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6241 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6301 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6361 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6421 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6481 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6541 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6601 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
