LOCUS       sequence-357795-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of SpEDC1opt_155-180.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  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 7049)
  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: pnEA-NvM-SpEdc1opt_155-180-GB1_AD
FEATURES             Location/Qualifiers
     source          1..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             88..1188
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="METEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             1204..1224
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1551..1569)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1565..1612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1787..1792,1793..1809,1810..1815))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1842..1860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1928..2032
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2033..2101,2102..2893)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2251..2270)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3064..3652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3553..3572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3806..3823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3838..3977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3903..3922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4063..4085
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4079..4270)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6625..6644
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6664..6741)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6947..6966
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
       61 tgtttaactt taagaaggag atataccatg gaaactgaag aaggtaaact ggtaatctgg
      121 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
      181 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
      241 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
      301 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
      361 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
      421 gaagcgttat cgctgattta taacaaagac ctgctgccga acccgccaaa aacctgggaa
      481 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgccct gatgttcaac
      541 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
      601 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
      661 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
      721 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
      781 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
      841 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
      901 ccgaacaaag agctggcgaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
      961 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     1021 ttggcgaaag atccacgtat tgccgccacg atggaaaacg cccagaaagg tgaaatcatg
     1081 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     1141 gccagcggtc gtcagactgt cgatgaagcc cttaaagacg cgcagactgg taccggcagc
     1201 ggcgaaaacc tttacttcca gggatcctcc attctttacg ccggtcctac atttactcat
     1261 tctccggccg cttctaatct tcccatccct acgttcttac actctctcga gatgcagtat
     1321 aaactgattc tgaatggcaa aaccctgaaa ggtgaaacca ccaccgaagc agttgatgca
     1381 gccaccgcag aaaaagtttt taaacagtat gccaatgata acggcgttga tggtgaatgg
     1441 acctatgatg atgcaaccaa aacctatacc gttaccgaat gatctagagc tagccctagg
     1501 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1561 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1621 aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     1681 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     1741 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     1801 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt
     1861 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1921 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1981 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2041 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2101 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2161 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2221 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     2281 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2341 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2401 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2461 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2521 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     2581 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2641 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2701 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2761 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2821 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2881 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2941 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3001 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3061 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3121 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3181 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3241 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3301 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3361 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3421 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3481 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3541 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3601 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3661 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3721 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3781 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3841 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     3901 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3961 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4021 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4081 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     4141 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     4201 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     4261 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     4321 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     4381 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     4441 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     4501 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     4561 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     4621 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     4681 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc
     4741 aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg
     4801 atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc
     4861 caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt
     4921 ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc
     4981 tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat
     5041 cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc
     5101 ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc
     5161 gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag
     5221 caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa
     5281 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     5341 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     5401 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     5461 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     5521 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     5581 ctcactgccc gctttccagt cgggaaactg tcgtgccagc tgcattaatg aatcggccaa
     5641 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     5701 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     5761 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     5821 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     5881 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5941 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     6001 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     6061 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     6121 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     6181 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     6241 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     6301 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     6361 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     6421 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     6481 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     6541 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     6601 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     6661 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     6721 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6781 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6841 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6901 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6961 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     7021 agaggatcga gatctcgatc ccgcgaaat
//
