LOCUS       sequence-274572-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector TMK1 X035_pECIA14 should be used with bait vector TMK1 
            X035_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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: TMK1 X035_pECIA14
FEATURES             Location/Qualifiers
     source          1..6584
                     /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             1880..1903
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3578..3601
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3617..3634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3657..3674)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3707..3841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3731..3750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3785..3804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3985..4001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3985..4001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3998..4020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4009..4025
                     /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(4033..4039,4040..4057,4058..4063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4078..4099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4216..4233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4387..4975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4467..4486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5146..5937,5938..6006))
                     /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     5769..5788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6007..6111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6179..6197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6235..6257)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6357..6376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6570..6584,1..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 tctgatggag atctctcagc
      541 gatgttatca ctcaagaaaa gcttaaaccc acccagttct ttcggttggt ctgaccctga
      601 cccatgtaaa tggactcaca tcgtttgtac aggaacaaaa cgtgtgaccc ggatccaaat
      661 cgggcattcg ggtcttcaag gtacactttc tcctgatcta cgtaacttat cagagcttga
      721 aagacttgag cttcaatgga acaacatctc tggtcctgtt ccttctttaa gtggtttagc
      781 ttcattacaa gtcttgatgt taagtaacaa caactttgat tctatcccta gtgatgtctt
      841 tcaaggttta acttcgttac aatctgtaga aattgataac aatcctttta agagttggga
      901 gattccagag agtttgagaa atgcttctgc tcttcagaat ttctctgcta attcggctaa
      961 tgtttccggt tctttacccg gttttctcgg accggatgag tttcccggtt tgtcgatttt
     1021 gcatttggct ttcaacaact tagaaggaga gttgcctatg agtttagctg gctctcaggt
     1081 tcagtcattg tggctcaatg gtcagaaact aaccggtgat attactgttc ttcagaacat
     1141 gactggttta aaagaggttt ggcttcactc caacaagttt tcgggtcctt taccggactt
     1201 ttcgggtctt aaggagcttg agagcttgag tttgagagat aactccttta caggtccagt
     1261 tcctgcgtct ttgttaagtc ttgagtcact taaagttgtg aacttgacga ataatcatct
     1321 acaaggacca gtgcctgtgt ttaagagctc tgtttcagtt gatttggata aagattctaa
     1381 cagcttttgc ttgtctagtc ctggtgagtg tgatcctaga gtgaagtctt tgcttttgat
     1441 agctagttca ttcgattatc cgccccggct cgctgagagt tggaaaggaa acgatccttg
     1501 tactaactgg attgggatag cttgtagcaa tgggaacatt actgttatca gtcttgagaa
     1561 aatggaacta accgggacga tttctcccga gtttggagcg atcaagtcgc ttcaaagaat
     1621 cattcttggt atcaacaatc ttactggtat gattcctcaa gagcttacaa cgttacctaa
     1681 tctcaaaaca ctcgatgttt cgagtaacaa gcttttcggg aaggtcccgg gttttagaag
     1741 caatgttgtt gtgaatacta atggtaatcc tgacattgga aaggataaaa gctctttgtc
     1801 ttctcctggt tcttcttcgc cttcaggtgg ttcgggttca ggtatcaacg gtgataaaga
     1861 ccggagagga atgaagtctc tggaggtgct gttccaggga ccaagcggca atcccggcgg
     1921 cgatctggcc ccccagatgc tgcgcgagct gcaggagacc aatgccgccc tgcaggacgt
     1981 gcgcgagctg ctgcgccacc gcgtgaagga gatcaccttc ctgaagaaca ccgtgatgga
     2041 gtgcgacgcc tgcggcatgc agccctctag aggctccggc tccatcatcc cagttgagga
     2101 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     2161 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     2221 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     2281 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     2341 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     2401 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     2461 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     2521 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     2581 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     2641 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtggag gccgaaagta
     2701 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     2761 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     2821 tgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     2881 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     2941 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     3001 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     3061 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     3121 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     3181 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     3241 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     3301 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     3361 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     3421 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     3481 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     3541 gcacccgggt tctagaggaa gcacctccgg cgataacgat tacaaggatg atgatgataa
     3601 ggccagcggc accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3661 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3721 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3781 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3841 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3901 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     3961 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4021 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4081 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4141 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6121 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6181 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6241 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6301 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6361 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6421 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6481 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6541 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
