LOCUS       sequence-419243-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nonsense mistranslation, TAA stop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5632)
  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 5632)
  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 TAA
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..144
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(203..994,995..1063))
                     /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     826..845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1064..1168)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1220..1247)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1455..1460,1461..1477,1478..1483)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1583..1611)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2206..2226
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2287..2988
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKG*SEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFED
                     GGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGD
                     IKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEG
                     RHSTGGMDELYK"
     primer_bind     complement(2426..2444)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2694..2713
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3091..3114
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3199..3293
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3785..4007
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4055..5005
                     /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(5543..5629)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctctagagc ttgcatgccc aaggagatgg cgcccaacag tcccccggcc acggggcctg
       61 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
      121 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
      181 ccacgatgcg tccggcgtag agttaccaat gcttaatcag tgaggcacct atctcagcga
      241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
      301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
      361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
      421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
      481 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
      541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
      601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
      661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
      721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
      781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
      841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
      901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
      961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     1081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     1201 aagaaaccat tattatcata aaaggccatc cgtcaggatg gccttctatg aatgccttgg
     1261 ctgcgattca ttctttatat gaataaaatt gttgtcagtt tcacgtcttg tcctgccata
     1321 tcgcgaaatt tctgcgcaaa agcacaaaaa atttttgcat ctcccccttg atgacgtggg
     1381 ttacgacccc atttagtagt caaccgcagt gagtgagtct gacgaggccc tttcgtcttc
     1441 aagaattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag
     1501 ttaaattgct aacgcagtcc ggggaattca ggcttggagg aacaatatga aaggatccaa
     1561 aggtgaagaa ttattcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1621 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1681 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
     1741 tttcgcgtat ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt
     1801 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     1861 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     1921 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
     1981 caactataac tcacacaaag tatacatcat ggcagacaaa caaaaggatg gaatcaaagt
     2041 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     2101 acgaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2161 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     2221 tgtaacagct gctgggatta cacatggtat ggatgaattg tacaaataac cgccaggagg
     2281 aacaatatgg tgagcaaggg ctaaagtgag gcagtgatca aggagttcat gcggttcaag
     2341 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     2401 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     2461 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac
     2521 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc
     2581 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac
     2641 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta
     2701 atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc
     2761 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg
     2821 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc
     2881 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa
     2941 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtg accgcctaat
     3001 gagcgggctt taatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc
     3061 tccgtcgaca agcttgcggc cgcactcgag caccaccacc accaccacca ccactaattg
     3121 attaatacct aggctgctaa acaaagccca tggatcgata gctggtcgac aggactctgg
     3181 ggttcgagag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     3241 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     3301 ctagggacag taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag
     3361 ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc
     3421 aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc
     3481 cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat
     3541 aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac
     3601 tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt
     3661 cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt
     3721 ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag
     3781 tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat
     3841 tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt
     3901 tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat
     3961 acccacaact caaaggaaaa ggactagtaa ttatcattga ctagcccatc tcaattggta
     4021 tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc
     4081 aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg
     4141 ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc
     4201 gttcacttat aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt
     4261 attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt
     4321 taaaggcttt gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga
     4381 attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata
     4441 taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt
     4501 attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga
     4561 atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg
     4621 ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga
     4681 taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga
     4741 tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac
     4801 cattacatca gattcctacc tacataacgg actaagaaaa acactacacg atgctttaac
     4861 tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagta
     4921 tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat
     4981 actggctaaa tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat
     5041 caagactaac aaacaaaagt agaacaactg ttcaccgtta catatcaaag ggaaaactgt
     5101 ccatatgcac agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt
     5161 ttggtgcatt caaagctgtt caccatgaac agatcgacaa tgtaacagat gaacagcatg
     5221 taacacctaa tagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac
     5281 acctgagaca acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc
     5341 tgcttatcga atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa
     5401 aatcatggca attctggaag aaatagcgct ttcagccggc aaaccggctg aagccggatc
     5461 tgcgattctg ataacaaact agcaacacca gaacagcccg tttgcgggca gcaaaacccg
     5521 taccctaggt ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     5581 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tg
//
