LOCUS       sequence-291279-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmBam.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  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 6791)
  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_AG
FEATURES             Location/Qualifiers
     source          1..6791
                     /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)"
     CDS             2489..3589
                     /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             3605..3625
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5023..5041)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5037..5084
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5259..5264,5265..5281,5282..5287))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5399..5944
                     /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     5845..5864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6061..6078
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6470..6572
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6573..6791,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6639..6658)
                     /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 gatataccat ggaaactgaa gaaggtaaac tggtaatctg
     2521 gattaacggc gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga
     2581 taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt
     2641 tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta
     2701 cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta
     2761 tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt
     2821 tgaagcgtta tcgctgattt ataacaaaga cctgctgccg aacccgccaa aaacctggga
     2881 agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgccc tgatgttcaa
     2941 cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa
     3001 gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc
     3061 gggtctgacc ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta
     3121 ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg
     3181 ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt
     3241 caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag
     3301 tccgaacaaa gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct
     3361 ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga
     3421 gttggcgaaa gatccacgta ttgccgccac gatggaaaac gcccagaaag gtgaaatcat
     3481 gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc
     3541 cgccagcggt cgtcagactg tcgatgaagc ccttaaagac gcgcagactg gtaccggcag
     3601 cggcgaaaac ctttacttcc agggccatat gctcgagatg cttaatgcac gtgacgtgtg
     3661 tcctgagggc aacgacgacc agcagttgga ccacaatttt aagcagatgg aggagcattt
     3721 ggccttaatg gtggaaggca atgaaaacga agatccgagg aaagccactt gtgagtacga
     3781 ggatacgaac gaagatggtg caacctgcac atcgggcgtt ttatccgaaa tccaggagaa
     3841 cttcggtaga ctccggttgt gtgacgttac tgcaccactc ctcgaattcc acggtttgga
     3901 ttgcttgcaa cagattcaaa agcgctcgcg ccattttgca ttcgacggtt ctccggccaa
     3961 gaagtcgcga tccggaggcg tgttggtcac cgggccaaag cagaagcaac tgcagaagga
     4021 aaatgtgtgg aaccggaaga gtaaaggctc tgcgtccgcg gataatattg agaaactgcc
     4081 cataactatt gagaaactgc atatgattgg tctgcacggc gattgcttag agcacaacgc
     4141 cgtgctgcgt ttgatgaatc tgttcagatc cctgcatgat cacctgaccg ccgatttggg
     4201 cttctcgcgc caaaactcaa tgccctcgga ctatctgttc gatatgccgg tgaagagcac
     4261 gatgcctaag agcttgaatg tgcgctacca actgcaggtg ctgtgcacca aagtagagcg
     4321 cttccttgtc cagcagcgcc gcaccttgga gtcgaatcgc cacttcgatt tcgagaaata
     4381 cgacgagtgt gacaagttgc ttaagggttt cgcatcctat ttggacaact tcaaactgct
     4441 tttaaagccc aaaatgcgca atcgaaacgg aaactcgggg agcaatgcgg acaagttcca
     4501 tactcagcgc atggagagat tgctaattgg tctgcgcgat tggatcaagg ctgcgcatct
     4561 cagtgtgcac gtatttaact gggaaatgga tctggagcac cgctactccg gggccatgac
     4621 cgaaagccac aagtcgttga acgagcgggc catccttttg tccggtgccg agctaagggc
     4681 ggccgaagcg cgtggaatca gtgcggagga tctgttcatc gcccagagat acaaactggg
     4741 aggtccgatc tattgcgttc tggagcagca tgagttcctc tccgctctga tcgccaatcc
     4801 agagacctat ttcccgccca gtgttgtcgc catttgcggg ccacagaagc ttggcgcagt
     4861 gagcatggag cagccgtcag cgtcggagga ggagtttgag gagaccgagg aagtgccatc
     4921 atcgccacct cgtcacaccg gacgtgtacc tcgcttcaga agctaaccta ggagatccgg
     4981 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     5041 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     5101 tatccggata tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     5161 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     5221 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag
     5281 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     5341 aagagataaa ttgcactgaa atcaagaaat attttatctg attaataaga tgatcttctt
     5401 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     5461 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     5521 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     5581 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     5641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     5701 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     5761 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     5821 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5881 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     5941 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     6001 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     6061 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     6121 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     6181 catagtaagc cagtatacac tcccctagcg ctgatgtccg gcggtgcttt tgccgttacg
     6241 caccaccccg tcagtagctg aacaggaggg acagctgata gaaacagaag ccactggagc
     6301 acctcaaaaa caccatcata cactaaatca gtaagttggc agcatcaccc gacgcacttt
     6361 gcgccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtgtcc
     6421 ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt gatcggcacg
     6481 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     6541 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     6601 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     6661 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     6721 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     6781 atccggaatt c
//
