LOCUS       sequence-290995-        5067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of SpDcp2opt_1-242.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Valkov E, Muthukumar S, Chang CT, Jonas S, Weichenrieder O, 
            Izaurralde E
  TITLE     Structure of the Dcp2-Dcp1 mRNA-decapping complex in the activated 
            conformation.
  JOURNAL   Nat Struct Mol Biol. 2016 Jun;23(6):574-9. doi: 10.1038/nsmb.3232. 
            Epub 2016 May 16.
  PUBMED    27183195
REFERENCE   2  (bases 1 to 5067)
  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/nsmb.3232";
            journalName: "Nat Struct Mol Biol"; date: "2016-06"; volume: "23"; 
            issue: "6"; pages: "574-9"
COMMENT     Sequence Label: pnYC-SpDcp2opt_1-242_AD
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2402..2421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2421..2445
                     /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)."
     RBS             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3299..3317)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3313..3360
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3535..3540,3541..3557,3558..3563))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3675..4220
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4121..4140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4337..4354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4746..4848
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4849..5067,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4915..4934)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatcagact aagatctcga tcccgcgaaa
     2401 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccact agtaataatt
     2461 ttgtttaact ttaagaagga gatatacata tgagctttac caatgcaacc tttagccagg
     2521 ttctggatga tctgagcgca cgttttattc tgaatctgcc tgcagaagaa cagagcagcg
     2581 ttgaacgtct gtgttttcag attgaacagg cccattggtt ttatgaggat tttattcgtg
     2641 cacagaatga tcagctgccg agcctgggtc tgcgtgtttt tagcgcaaaa ctgtttgcac
     2701 attgtccgct gctgtggaaa tggtcaaaag ttcatgaaga ggcctttgat gatttcctgc
     2761 gttataaaac ccgtattccg gttcgtggtg caattatgct ggatatgagc atgcagcagt
     2821 gtgttctggt gaaaggttgg aaagcaagca gcggttgggg ttttccgaaa ggtaaaattg
     2881 ataaagatga gagcgacgtg gattgtgcaa ttcgtgaagt ttatgaagaa accggctttg
     2941 attgtagcag ccgtattaat ccgaacgagt ttattgatat gaccattcgt ggtcagaatg
     3001 tgcgcctgta tatcattccg ggtattagcc tggatacccg ttttgaaagc cgtacccgta
     3061 aagaaattag caaaatcgaa tggcacaacc tgatggatct gccgaccttc aaaaaaaaca
     3121 aaccgcagac catgaaaaac aaattctata tggtgatccc gtttctggca ccgctgaaaa
     3181 aatggatcaa aaaacgtaac attgccaaca acacctaggg atcctaatag tctagagcta
     3241 gccctaggag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     3301 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3361 ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca
     3421 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
     3481 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     3541 gcttatcgat gataagctgt caaacatgag aattacaact tatatcgtat ggggctgact
     3601 tcaggtgcta catttgaaga gataaattgc actgaaatca agaaatattt tatctgatta
     3661 ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa
     3721 aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga
     3781 ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg
     3841 gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc
     3901 ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3961 ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact
     4021 gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc
     4081 gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt
     4141 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     4201 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat
     4261 cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag
     4321 tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat
     4381 attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca
     4441 ccctcatcag tgccaacata gtaagccagt atacactccc ctagcgctga tgtccggcgg
     4501 tgcttttgcc gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa
     4561 cagaagccac tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca
     4621 tcacccgacg cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa
     4681 ataaatcctg gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga
     4741 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     4801 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     4861 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     4921 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     4981 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     5041 cgcctgatga atgctcatcc ggaattc
//
