LOCUS       sequence-291080-        7346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsRoquin1_510-1133.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7346)
  AUTHORS   Sgromo A, Raisch T, Bawankar P, Bhandari D, Chen Y, Kuzuoglu-Ozturk 
            D, Weichenrieder O, Izaurralde E
  TITLE     A CAF40-binding motif facilitates recruitment of the CCR4-NOT 
            complex to mRNAs targeted by Drosophila Roquin.
  JOURNAL   Nat Commun. 2017 Feb 6;8:14307. doi: 10.1038/ncomms14307.
  PUBMED    28165457
REFERENCE   2  (bases 1 to 7346)
  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"; journalName: "Nat 
            Commun."; date: "2017-02-6"; volume: "8:14307. doi"; pages: " 
            10.1038/ncomms14307"
COMMENT     Sequence Label: pnYC-NpM-HsRoquin1_510-1133_AE
FEATURES             Location/Qualifiers
     source          1..7346
                     /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..3628
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             4324..4332
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(5578..5596)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5592..5639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5814..5819,5820..5836,5837..5842))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5954..6499
                     /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     6400..6419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6616..6633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7025..7127
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(7128..7346,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7194..7213)
                     /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 cggcctggaa gttctgttcc aggggcccca tatgctcgag cttaagcgag ggacagaccc
     3661 cagctatgat tctagtctga aaccaggaaa aatagatcat ctgagcagta gtgctcctgg
     3721 atcccctcct gacctgctag aatctgttcc taagagtatt tctgccttac ctgtgaatcc
     3781 acattctata cctccaaggg ggccagcaga tctgcctcca atgcctgtta ccaaaccact
     3841 tcagatggta cctcgaggtt ctcagttata tccagcacaa cagacggatg tttattatca
     3901 ggatcctcga ggagcagctc cgccatttga accagcacct tatcagcagg gtatgtatta
     3961 tactccacca ccacaatgtg tgtcccgctt tgtccgacct ccaccatctg ctcctgaacc
     4021 tgctcctccc tacttggatc attatccacc ctacctccaa gaacgtgttg taaactctca
     4081 gtatggcaca cagccacagc agtacccacc tatataccca tctcactatg atggccgtcg
     4141 agtgtaccct gctccgtctt acacaagaga agagatattc cgagaaagcc ctatacccat
     4201 tgagattcca cctgcagcag taccatcgta tgtaccagaa tccagagaaa gataccaaca
     4261 gatcgagagt tactatccag tggctcctca tccaactcag atcagacctt cgtacctcag
     4321 agaacctcct tatagccggc ttcctcctcc tccccagcct catcctagtc tagatgaact
     4381 acatcgccga cgaaaggaaa taatggccca gctagaggaa agaaaggtta tctctccacc
     4441 tccttttgca ccttcaccaa ccttgcctcc tacctttcat ccggaagaat ttttggatga
     4501 agacttgaag gtagctggga aatacaaagg aaatgattat agccaatact ctccctggtc
     4561 atgtgacacc atcggctcct acattggaac caaagatgca aaacccaaag atgttgtggc
     4621 agcagggagt gtggaaatga tgaatgtgga gagtaaagga atgagggacc agcgattaga
     4681 tcttcagaga agagcagcag aaaccagtga tgatgacctc atcccatttg gagaccgacc
     4741 aacagtgtct cggtttggtg ccatctcacg aacttccaaa actatatatc agggtgctgg
     4801 tccaatgcag gctatggcac ctcagggagc tcctacaaaa tctattaaca tttcagatta
     4861 tagtccatat ggaacccacg gtggctgggg agcttctcca tattcacctc atcaaaacat
     4921 accttctcag ggacacttca gtgagaggga gagaatatct atgtcagaag tggccagtca
     4981 tggaaaaccc cttccatctg ctgagagaga acagctacga ctagaattgc agcaattgaa
     5041 ccatcagatt agccagcaga cccagctacg tggactagag gctgttagta acaggctggt
     5101 gttgcagagg gaggcaaaca ccctggcagg ccagtcacag cccccaccac cgccacctcc
     5161 aaaatggcct gggatgatct caagtgagca gttgagcttg gaactgcacc aggtggaaag
     5221 ggaaatcggg aagagaacac gggaactgag tatggaaaac cagtgttctc tggacatgaa
     5281 aagcaaactg aatacaagta aacaagcaga aaatggacaa ccagaaccac aaaacaaggt
     5341 tccggctgag gaccttacat tgacattcag tgatgtacca aatggatcag ccttgacaca
     5401 agagaatatc agcctcctat caaacaagac cagctctctg aacctgtcag aggaccctga
     5461 gggaggaggg gataataatg actcccagag atcaggagtt actcccagtt ctgctcctta
     5521 acctaggaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     5581 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     5641 tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa
     5701 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     5761 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     5821 cttatcgatg ataagctgtc aaacatgaga attacaactt atatcgtatg gggctgactt
     5881 caggtgctac atttgaagag ataaattgca ctgaaatcaa gaaatatttt atctgattaa
     5941 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     6001 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     6061 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     6121 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     6181 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6241 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     6301 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     6361 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     6421 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     6481 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     6541 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     6601 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     6661 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     6721 cctcatcagt gccaacatag taagccagta tacactcccc tagcgctgat gtccggcggt
     6781 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac
     6841 agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat
     6901 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa
     6961 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag
     7021 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     7081 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     7141 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     7201 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     7261 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     7321 gcctgatgaa tgctcatccg gaattc
//
