LOCUS       sequence-356589-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmNanos-opt_116-163-F152E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  AUTHORS   Raisch T, Bhandari D, Sabath K, Helms S, Valkov E, Weichenrieder O, 
            Izaurralde E
  TITLE     Distinct modes of recruitment of the CCR4-NOT complex by Drosophila 
            and vertebrate Nanos.
  JOURNAL   EMBO J. 2016 May 2;35(9):974-90. doi: 10.15252/embj.201593634. Epub 
            2016 Mar 11.
  PUBMED    26968986
REFERENCE   2  (bases 1 to 6524)
  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: 
            "10.15252/embj.201593634"; journalName: "EMBO J"; date: "2016-05-2- 
            2"; volume: "35"; issue: "9"; pages: "974-90"
COMMENT     Sequence Label: pnEA-NpG-DmNanos-opt_116-163-F152E_AB
FEATURES             Location/Qualifiers
     source          1..6524
                     /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..741
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             763..786
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1026..1044)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1040..1087
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1262..1267,1268..1284,1285..1290))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1317..1335)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1403..1507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1508..1576,1577..2368)
                     /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(1726..1745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2539..3127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3028..3047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3281..3298
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3313..3452
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3378..3397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3538..3560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3554..3745)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6100..6119
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6139..6216)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6422..6441
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
       61 tgtttaactt taagaaggag atataccatg gcccctatac taggttattg gaaaattaag
      121 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
      181 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
      241 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
      301 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
      361 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
      421 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
      481 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
      541 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
      601 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
      661 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
      721 ggtggcgacc atccgccaaa atcggatggt accggcagcg gcctggaagt tctgttccag
      781 gggccccata tgctcgagtc acatcagcag accgatgaaa ttgcacgtag cctgaaaatc
      841 tttgcacagg ttaccaccgg tgcagcagaa aatgcagcag gtagcatgca ggatgttatg
      901 caagaagaag caaccaatgg ttatgccagt gatgatctgg gttaaggatc ctaatagtct
      961 agagctagcc ctaggagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1021 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1081 ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc
     1141 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca
     1201 ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg
     1261 cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct
     1321 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1381 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1441 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1501 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1561 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1621 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1681 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1741 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1801 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1861 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1921 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1981 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2041 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2101 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2161 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2221 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2281 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2341 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2401 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2461 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2521 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2581 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2641 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2701 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2761 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2821 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2881 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2941 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3001 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3061 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3121 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3181 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3241 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3301 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3361 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3421 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3481 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3541 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3601 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3661 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3721 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3781 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     3841 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     3901 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     3961 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4021 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4081 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     4141 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     4201 tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga
     4261 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca
     4321 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc
     4381 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     4441 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata
     4501 aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga
     4561 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     4621 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     4681 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     4741 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     4801 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     4861 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     4921 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     4981 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aactgtcgtg ccagctgcat
     5101 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5161 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5221 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5281 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5341 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5401 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5461 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5521 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5581 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5641 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5701 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5761 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5821 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5881 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5941 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6001 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6061 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6121 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6181 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6241 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6301 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6361 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6421 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6481 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
