LOCUS       sequence-357802-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmBam_141-422.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  AUTHORS   Sgromo A, Raisch T, Backhaus C, Keskeny C, Alva V, Weichenrieder O, 
            Izaurralde E
  TITLE     Drosophila Bag-of-marbles directly interacts with the CAF40 subunit 
            of the CCR4-NOT complex to elicit repression of mRNA targets.
  JOURNAL   RNA. 2018 Mar;24(3):381-395. doi: 10.1261/rna.064584.117. Epub 2017 
            Dec 18.
  PUBMED    29255063
REFERENCE   2  (bases 1 to 5930)
  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: 
            "10.1261/rna.064584.117"; journalName: "RNA"; date: "2018-03"; 
            volume: "24"; issue: "3"; pages: "381-395"
COMMENT     Sequence Label: pnYC-NvM-DmBam_141-442_AG
FEATURES             Location/Qualifiers
     source          1..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1825..1844
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1933..1952
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1933..1951
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1952..1976
                     /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             1991..2013
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2020..3120
                     /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             3136..3156
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4134..4152)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4148..4195
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4370..4375,4376..4392,4393..4398))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4510..5055
                     /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     4956..4975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5172..5189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5581..5683
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5684..5930,1..413)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5750..5769)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
       61 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
      121 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
      181 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
      241 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
      301 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
      361 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
      421 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
      481 ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg
      541 ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt
      601 gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat
      661 aattgacgat atgatcattt attctgcctc ccagagcctg ataaaaacgg ttagcgcttc
      721 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
      781 aacataatgg tgcagggcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccagg
      841 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
      901 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
      961 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1021 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     1081 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     1141 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     1201 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     1261 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     1321 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     1381 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1441 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1501 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1561 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1621 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     1681 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     1741 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     1801 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     1861 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcagac taagatctcg
     1921 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccac
     1981 tagtaataat tttgtttaac tttaagaagg agatatacca tggaaactga agaaggtaaa
     2041 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
     2101 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
     2161 ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
     2221 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
     2281 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
     2341 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag acctgctgcc gaacccgcca
     2401 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcc
     2461 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
     2521 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
     2581 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
     2641 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
     2701 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
     2761 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
     2821 aacgccgcca gtccgaacaa agagctggcg aaagagttcc tcgaaaacta tctgctgact
     2881 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     2941 tacgaggaag agttggcgaa agatccacgt attgccgcca cgatggaaaa cgcccagaaa
     3001 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     3061 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag cccttaaaga cgcgcagact
     3121 ggtaccggca gcggtgaaaa cctttacttc cagggccata tgctcgaggc ggataatatt
     3181 gagaaactgc ccataactat tgagaaactg catatgattg gtctgcacgg cgattgctta
     3241 gagcacaacg ccgtgctgcg tttgatgaat ctgttcagat ccctgcatga tcacctgacc
     3301 gccgatttgg gcttctcgcg ccaaaactca atgccctcgg actatctgtt cgatatgccg
     3361 gtgaagagca cgatgcctaa gagcttgaat gtgcgctacc aactgcaggt gctgtgcacc
     3421 aaagtagagc gcttccttgt ccagcagcgc cgcaccttgg agtcgaatcg ccacttcgat
     3481 ttcgagaaat acgacgagtg tgacaagttg cttaagggtt tcgcatccta tttggacaac
     3541 ttcaaactgc ttttaaagcc caaaatgcgc aatcgaaacg gaaactcggg gagcaatgcg
     3601 gacaagttcc atactcagcg catggagaga ttgctaattg gtctgcgcga ttggatcaag
     3661 gctgcgcatc tcagtgtgca cgtatttaac tgggaaatgg atctggagca ccgctactcc
     3721 ggggccatga ccgaaagcca caagtcgttg aacgagcggg ccatcctttt gtccggtgcc
     3781 gagctaaggg cggccgaagc gcgtggaatc agtgcggagg atctgttcat cgcccagaga
     3841 tacaaactgg gaggtccgat ctattgcgtt ctggagcagc atgagttcct ctccgctctg
     3901 atcgccaatc cagagaccta tttcccgccc agtgttgtcg ccatttgcgg gccacagaag
     3961 cttggcgcag tgagcatgga gcagccgtca gcgtcggagg aggagtttga ggagaccgag
     4021 gaagtgccat catcgccacc tcgtcacacc ggacgtgtac ctcgcttcag aagctaacct
     4081 aggagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     4141 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     4201 aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     4261 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     4321 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     4381 tcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg
     4441 tgctacattt gaagagataa attgcactga aatcaagaaa tattttatct gattaataag
     4501 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
     4561 cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
     4621 ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
     4681 tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
     4741 catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
     4801 gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
     4861 tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
     4921 gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt
     4981 cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg
     5041 cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc
     5101 tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa
     5161 cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct
     5221 gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc
     5281 atcagtgcca acatagtaag ccagtataca ctcccctagc gctgatgtcc ggcggtgctt
     5341 ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa
     5401 gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc
     5461 cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
     5521 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt
     5581 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     5641 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     5701 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     5761 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     5821 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     5881 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
//
