LOCUS       sequence-275960-        6512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G24230 X040_pECIA14 should be used with bait vector 
            AT2G24230 X040_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6512)
  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 6512)
  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: AT2G24230 X040_pECIA14
FEATURES             Location/Qualifiers
     source          1..6512
                     /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             1808..1831
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3506..3529
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3545..3562
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3585..3602)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3635..3769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3713..3732
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3913..3929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3913..3929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3926..3948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3937..3953
                     /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(3961..3967,3968..3985,3986..3991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4006..4027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4144..4161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4315..4903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4395..4414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5074..5865,5866..5934))
                     /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     5697..5716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5935..6039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6107..6125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6163..6185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6285..6304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6498..6512,1..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 aacacagatg gtttctttgt
      541 ctctgaattc tataagcaaa tgggtttaag ctcttctcaa gcttataact tctctgctcc
      601 tttttgttca tggcaaggtt tgttctgtga ttccaagaat gagcatgtga ttatgttaat
      661 tgcttctgga atgagtttat ctggtcaaat ccctgataac acaattggta aactcagcaa
      721 gcttcaatct ttggatctta gcaacaacaa aatctctgct ttaccatctg atttctggag
      781 tttgaacact ttgaagaatc tcaacctctc tttcaacaag atctctgggt ctttctctag
      841 taatgttggc aattttgggc agcttgagtt acttgatatc tcttacaaca atttctctgg
      901 tgcaatccct gaagctgtgg attctcttgt tagtttgaga gtcttgaagc ttgatcataa
      961 tgggtttcag atgtcgattc cacgaggact tcttggttgc caatccctgg tttctattga
     1021 tctctcctct aaccagcttg aagggtctct tcctgatggg tttggctctg catttccaaa
     1081 gcttgagact ttgagccttg caggaaacaa gattcatggc agggacacag attttgctga
     1141 tatgaaatcc atcagttttc tcaacatttc aggaaatcag tttgatggtt cagtgacagg
     1201 tgtgttcaag gagactttag aagtggctga tctcagcaaa aaccggtttc aaggtcatat
     1261 ctcatctcag gtagactcca actggtttag tttggtttat cttgacttgt ctgagaatga
     1321 gctcagtgga gttatcaaga atctgacact actaaagaag ctcaagcact taaatctagc
     1381 ttggaataga ttcaacagag gaatgtttcc gaggattgag atgctttctg gtttagagta
     1441 tcttaacttg tcaaacacaa atctttctgg tcacatacct agagagatct caaagctgag
     1501 tgatttgagt acactcgatg tttctggaaa ccatcttgca ggacatatcc cgattttgag
     1561 tatcaagaac cttgtggcta tcgatgtgtc gaggaacaat ttgacgggag agataccgat
     1621 gtctatcctt gagaagctcc cttggatgga gagattcaac ttctctttca acaacttaac
     1681 attttgttcc ggaaagttct cagctgaaac tctgaacaga tccttctttg gttctacaaa
     1741 tagctgccct atcgctgcaa acccagcact tttcaagaga aaaagatcag tcactggagg
     1801 acttaagctg gaggtgctgt tccagggacc aagcggcaat cccggcggcg atctggcccc
     1861 ccagatgctg cgcgagctgc aggagaccaa tgccgccctg caggacgtgc gcgagctgct
     1921 gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc gtgatggagt gcgacgcctg
     1981 cggcatgcag ccctctagag gctccggctc catcatccca gttgaggagg agaacccgga
     2041 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     2101 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     2161 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     2221 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     2281 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     2341 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     2401 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     2461 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     2521 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     2581 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     2641 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     2701 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     2761 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     2821 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     2881 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     2941 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     3001 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     3061 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     3121 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     3181 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     3241 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     3301 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     3361 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     3421 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggttc
     3481 tagaggaagc acctccggcg ataacgatta caaggatgat gatgataagg ccagcggcac
     3541 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3601 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     3661 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3721 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3781 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3841 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     3901 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3961 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4021 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6061 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6121 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6181 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6241 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6361 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6421 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6481 ggtaacgcca gggttttccc agtcacgacg tt
//
