LOCUS       sequence-396630-        3278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bicistronic RBS 25K Part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3278)
  AUTHORS   Jansen Z, Reilly SR, Lieber-Kotz M, Li AZ, Wei Q, Kulhanek DL, 
            Gilmour AR, Thyer R
  TITLE     Interrogating the Function of Bicistronic Translational Control 
            Elements to Improve Consistency of Gene Expression.
  JOURNAL   ACS Synth Biol. 2023 Jun 16;12(6):1608-1615. doi: 
            10.1021/acssynbio.3c00093. Epub 2023 May 30.
  PUBMED    37253269
REFERENCE   2  (bases 1 to 3278)
  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: 
            "10.1021/acssynbio.3c00093"; journalName: "ACS Synth Biol"; date: 
            "2023-06-16- 16"; volume: "12"; issue: "6"; pages: "1608-1615"
COMMENT     Sequence Label: pTHY_P0030_T2c_BCD_25K
FEATURES             Location/Qualifiers
     source          1..3278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        112..161
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        524..595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(596..664,665..1456)
                     /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(814..833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1680..1729
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1841..2146
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2025..2044
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      2406..2994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2895..2914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acagggctat aaggtagatc cgggtcgaca gatataatta gccaaaaatt tctgtaataa
       61 atatcactag ctcagattca gtagaccgct gttggatagg tctctggaga gctgtcaccg
      121 gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa ataattttgt
      181 ttaagggccc aagttcactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa
      241 catcttaatc atgcggtaag gagttaaata tgagagacca tcgtgcatga tctacgtgcg
      301 tcacatgcag tacctgaatt gactctcttc cggggatcaa atgtagcacc tgaagtcacc
      361 ctcggtacca aattccagaa aagaggcctc ccgaaagggg ggcctttttt cgttttggtc
      421 cgttactgga acgttgtgag gtacacggaa gctgtggtat ggctgtgcag gtcgtaaatc
      481 actgcataat tcgtgtcgct caaggcgcat atttttctaa atacattcaa atatgtatcc
      541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
      841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     1141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gatcgcgcgg
     1321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1441 gattaagcat tggtaactgt tagaccaagt ttactcatat atactttaga ttgatttaaa
     1501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1561 aatcccttaa cgtgagtttt cgttccactg agcgtcgagt ggctgtcgac cggcgaaaag
     1621 ttcgattaca gctagctcag tcctaggtac aattacagcc atcgtacgag cccggagagc
     1681 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat
     1741 aattttgttt aagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg
     1801 tactgaaaca tcttaatcat gcggattcta gttaaatatg atgcagttta aggtttacac
     1861 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     1921 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     1981 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     2041 tatggccagt gtgccggtgt ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     2101 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatcc gttttagttc
     2161 gtcggtcagt ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag
     2221 gggcagaaag atgaatgact gtcgctgtgt acgcgataga taaaaaaaaa ggtttagaca
     2281 gtgcgaaagc actgtctaaa cctttttttt cctgctcagg cgcaaccgat catgaccaaa
     2341 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2401 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2461 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2521 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2581 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2641 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2701 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2761 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2821 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2881 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2941 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3001 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3061 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3121 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcccctgttt cccgactgga
     3181 aagcggcgac tcctgcatta ggaaattata aacgcaggtc ttttcgccgg tgtatttctt
     3241 gcaaaaaaaa accccgccga agcggggttt tttttttt
//
