LOCUS       sequence-417833-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  frameshift mistranslation, A4.
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 A4
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(145..163)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     413..432
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             810..833
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      918..1012
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1504..1726
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1774..2724
                     /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(3262..3348)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3476..3495
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(3554..4345,4346..4414))
                     /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     4177..4196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4415..4519)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4571..4598)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(4806..4811,4812..4828,4829..4834)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4934..4962)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5557..5577
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaacaatatg gtgagcaagg gcaaaagagg cagtgatcaa ggagttcatg cggttcaagg
       61 tgcacatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      121 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      181 tctcctggga catcctgtcc cctcagttca tgtacggctc cagggccttc atcaagcacc
      241 ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg
      301 tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg
      361 gcaccctgat ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa
      421 tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg
      481 tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg
      541 acttcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg
      601 accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac
      661 gctccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga ccgcctaatg
      721 agcgggcttt aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct
      781 ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccaccac cactaattga
      841 ttaataccta ggctgctaaa caaagcccat ggatcgatag ctggtcgaca ggactctggg
      901 gttcgagagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
      961 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac
     1021 tagggacagt aagacgggta agcctgttga tgataccgct gccttactgg gtgcattagc
     1081 cagtctgaat gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca
     1141 ggaactgctg aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc
     1201 ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata
     1261 aaaggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact
     1321 gaatgtcacg aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc
     1381 agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt
     1441 tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt
     1501 agtgagttat acacagggct gggatctatt ctttttatct ttttttattc tttctttatt
     1561 ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt
     1621 acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata
     1681 cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat
     1741 agtgattaaa atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca
     1801 aatgaactag cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc
     1861 tgtgtggcac tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg
     1921 ttcacttata accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta
     1981 ttagctaaag caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt
     2041 aaaggctttg agattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa
     2101 ttagttttta gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat
     2161 aatctggaac atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta
     2221 ttaaaagaac taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa
     2281 tttaagttca tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg
     2341 gttttgaaac caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat
     2401 aagcgaggcc gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat
     2461 ctcgtaaccg aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc
     2521 attacatcag attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact
     2581 gcaaaaattc agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat
     2641 gatctcaatg gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata
     2701 ctggctaaat acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc
     2761 aagactaaca aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc
     2821 catatgcaca gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt
     2881 tggtgcattc aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt
     2941 aacacctaat agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca
     3001 cctgagacaa cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct
     3061 gcttatcgaa tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa
     3121 atcatggcaa ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct
     3181 gcgattctga taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt
     3241 accctaggtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     3301 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagag
     3361 cttgcatgcc caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     3421 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     3481 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     3541 gtccggcgta gagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3841 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     3901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4141 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     4201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4561 ttattatcat aaaaggccat ccgtcaggat ggccttctat gaatgccttg gctgcgattc
     4621 attctttata tgaataaaat tgttgtcagt ttcacgtctt gtcctgccat atcgcgaaat
     4681 ttctgcgcaa aagcacaaaa aatttttgca tctccccctt gatgacgtgg gttacgaccc
     4741 catttagtag tcaaccgcag tgagtgagtc tgacgaggcc ctttcgtctt caagaattct
     4801 catgtttgac agcttatcat cgataagctt taatgcggta gtttatcaca gttaaattgc
     4861 taacgcagtc cggggaattc aggcttggag gaacaatatg aaaggatcca aaggtgaaga
     4921 attattcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
     4981 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt
     5041 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttcgcgta
     5101 tggtcttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact ttttcaagag
     5161 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
     5221 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
     5281 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa
     5341 ctcacacaaa gtatacatca tggcagacaa acaaaaggat ggaatcaaag ttaacttcaa
     5401 aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc aacgaaatac
     5461 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc
     5521 cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc
     5581 tgctgggatt acacatggta tggatgaatt gtacaaataa ccgccaggag
//
