LOCUS       sequence-417989-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  frameshift mistranslation, C3T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  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 5630)
  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 C3T
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(195..986,987..1055))
                     /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     818..837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1056..1160)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1212..1239)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1447..1452,1453..1469,1470..1475)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1575..1603)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2198..2218
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2416..2434)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2684..2703
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3081..3104
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3189..3283
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3775..3997
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4045..4995
                     /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(5533..5619)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcttgcatgc ccaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
       61 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
      121 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
      181 cgtccggcgt agagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      241 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      301 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      361 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      421 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      481 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
      541 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      601 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
      661 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
      721 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
      781 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
      841 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
      901 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
      961 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1021 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1081 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1141 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     1201 attattatca taaaaggcca tccgtcagga tggccttcta tgaatgcctt ggctgcgatt
     1261 cattctttat atgaataaaa ttgttgtcag tttcacgtct tgtcctgcca tatcgcgaaa
     1321 tttctgcgca aaagcacaaa aaatttttgc atctccccct tgatgacgtg ggttacgacc
     1381 ccatttagta gtcaaccgca gtgagtgagt ctgacgaggc cctttcgtct tcaagaattc
     1441 tcatgtttga cagcttatca tcgataagct ttaatgcggt agtttatcac agttaaattg
     1501 ctaacgcagt ccggggaatt caggcttgga ggaacaatat gaaaggatcc aaaggtgaag
     1561 aattattcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     1621 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     1681 ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact actttcgcgt
     1741 atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac tttttcaaga
     1801 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     1861 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     1921 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
     1981 actcacacaa agtatacatc atggcagaca aacaaaagga tggaatcaaa gttaacttca
     2041 aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat caacgaaata
     2101 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg
     2161 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag
     2221 ctgctgggat tacacatggt atggatgaat tgtacaaata accgccagga ggaacaatat
     2281 ggtgagcaag ggcccctgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     2341 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2401 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     2461 acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca
     2521 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2581 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     2641 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     2701 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     2761 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     2821 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     2881 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     2941 gccgccactc caccggcggc atggacgagc tgtacaagtg accgcctaat gagcgggctt
     3001 taatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tccgtcgaca
     3061 agcttgcggc cgcactcgag caccaccacc accaccacca ccactaattg attaatacct
     3121 aggctgctaa acaaagccca tggatcgata gctggtcgac aggactctgg ggttcgagag
     3181 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     3241 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagggacag
     3301 taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa
     3361 tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct
     3421 gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc
     3481 ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat aaaaggcgcc
     3541 tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac
     3601 gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt cagcgatttg
     3661 cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt ttgtagagga
     3721 gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag tagtgagtta
     3781 tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat tctataaatt
     3841 ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt
     3901 ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat acccacaact
     3961 caaaggaaaa ggactagtaa ttatcattga ctagcccatc tcaattggta tagtgattaa
     4021 aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc aaatgaacta
     4081 gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg ctgtgtggca
     4141 ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc gttcacttat
     4201 aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt attagctaaa
     4261 gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt taaaggcttt
     4321 gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga attagttttt
     4381 agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata taatctggaa
     4441 catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt attaaaagaa
     4501 ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga atttaagttc
     4561 atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa
     4621 ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga taagcgaggc
     4681 cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga tctcgtaacc
     4741 gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac cattacatca
     4801 gattcctacc tacataacgg actaagaaaa acactacacg atgctttaac tgcaaaaatt
     4861 cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagta tgatctcaat
     4921 ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat actggctaaa
     4981 tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat caagactaac
     5041 aaacaaaagt agaacaactg ttcaccgtta catatcaaag ggaaaactgt ccatatgcac
     5101 agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt ttggtgcatt
     5161 caaagctgtt caccatgaac agatcgacaa tgtaacagat gaacagcatg taacacctaa
     5221 tagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac acctgagaca
     5281 acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc tgcttatcga
     5341 atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa aatcatggca
     5401 attctggaag aaatagcgct ttcagccggc aaaccggctg aagccggatc tgcgattctg
     5461 ataacaaact agcaacacca gaacagcccg tttgcgggca gcaaaacccg taccctaggt
     5521 ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     5581 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctaga
//
