LOCUS       sequence-419241-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nonsense mistranslation, TAG stop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  AUTHORS   Hinnu M, Putrins M, Kogermann K, Kaldalu N, Tenson T
  TITLE     Fluorescent reporters give new insights into antibiotics-induced 
            nonsense and frameshift mistranslation.
  JOURNAL   Sci Rep. 2024 Mar 22;14(1):6883. doi: 10.1038/s41598-024-57597-8.
  PUBMED    38519558
REFERENCE   2  (bases 1 to 5633)
  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: "Sci Rep."; 
            date: "2024-03-22"; volume: "14(1):6883. doi"; pages: " 
            10.1038/s41598-024-57597-8"
COMMENT     Sequence Label: pSC101-GFPmut2-mScarlet-I TAG
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(190..981,982..1050))
                     /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     813..832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1051..1155)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1207..1234)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1442..1447,1448..1464,1465..1470)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1570..1598)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2193..2213
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2414..2432)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2682..2701
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3079..3102
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3187..3281
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3773..3995
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4043..4993
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(5531..5617)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 catgcccaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
       61 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
      121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
      181 ggcgtagagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      481 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
      541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      781 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
      841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     1201 tatcataaaa ggccatccgt caggatggcc ttctatgaat gccttggctg cgattcattc
     1261 tttatatgaa taaaattgtt gtcagtttca cgtcttgtcc tgccatatcg cgaaatttct
     1321 gcgcaaaagc acaaaaaatt tttgcatctc ccccttgatg acgtgggtta cgaccccatt
     1381 tagtagtcaa ccgcagtgag tgagtctgac gaggcccttt cgtcttcaag aattctcatg
     1441 tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac
     1501 gcagtccggg gaattcaggc ttggaggaac aatatgaaag gatccaaagg tgaagaatta
     1561 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1621 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1681 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt cgcgtatggt
     1741 cttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc
     1801 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1861 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1921 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
     1981 cacaaagtat acatcatggc agacaaacaa aaggatggaa tcaaagttaa cttcaaaatt
     2041 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaacg aaatactcca
     2101 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     2161 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
     2221 gggattacac atggtatgga tgaattgtac aaataaccgc caggaggaac aatatggtga
     2281 gcaagggcta gagtggaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
     2341 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2401 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
     2461 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
     2521 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2581 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     2641 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
     2701 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     2761 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
     2821 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     2881 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
     2941 cgccactcca ccggcggcat ggacgagctg tacaagtgac cgcctaatga gcgggcttta
     3001 atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc cgtcgacaag
     3061 cttgcggccg cactcgagca ccaccaccac caccaccacc actaattgat taatacctag
     3121 gctgctaaac aaagcccatg gatcgatagc tggtcgacag gactctgggg ttcgagagct
     3181 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     3241 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact agggacagta
     3301 agacgggtaa gcctgttgat gataccgctg ccttactggg tgcattagcc agtctgaatg
     3361 acctgtcacg ggataatccg aagtggtcag actggaaaat cagagggcag gaactgctga
     3421 acagcaaaaa gtcagatagc accacatagc agacccgcca taaaacgccc tgagaagccc
     3481 gtgacgggct tttcttgtat tatgggtagt ttccttgcat gaatccataa aaggcgcctg
     3541 tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga
     3601 aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgatttgcc
     3661 cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc
     3721 aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata
     3781 cacagggctg ggatctattc tttttatctt tttttattct ttctttattc tataaattat
     3841 aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct
     3901 agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca
     3961 aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata gtgattaaaa
     4021 tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc
     4081 gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact
     4141 actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa
     4201 ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc
     4261 aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggctttga
     4321 gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag
     4381 tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca
     4441 tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact
     4501 aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat
     4561 gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc
     4621 aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg
     4681 cccgactgat acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga
     4741 acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga
     4801 ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg caaaaattca
     4861 gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg atctcaatgg
     4921 ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata
     4981 cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa
     5041 acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc atatgcacag
     5101 atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt ggtgcattca
     5161 aagctgttca ccatgaacag atcgacaatg taacagatga acagcatgta acacctaata
     5221 gaacaggtga aaccagtaaa acaaagcaac tagaacatga aattgaacac ctgagacaac
     5281 ttgttacagc tcaacagtca cacatagaca gcctgaaaca ggcgatgctg cttatcgaat
     5341 caaagctgcc gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa tcatggcaat
     5401 tctggaagaa atagcgcttt cagccggcaa accggctgaa gccggatctg cgattctgat
     5461 aacaaactag caacaccaga acagcccgtt tgcgggcagc aaaacccgta ccctaggtct
     5521 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     5581 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagagc ttg
//
