LOCUS       sequence-417985-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  frameshift mistranslation, A5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  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 5631)
  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 A5
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..146
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(205..996,997..1065))
                     /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     828..847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1066..1170)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1222..1249)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1457..1462,1463..1479,1480..1485)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1585..1613)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2208..2228
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2427..2445)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2695..2714
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3092..3115
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3200..3294
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3786..4008
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4056..5006
                     /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(5544..5630)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcctctaga gcttgcatgc ccaaggagat ggcgcccaac agtcccccgg ccacggggcc
       61 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
      121 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
      181 ggccacgatg cgtccggcgt agagttacca atgcttaatc agtgaggcac ctatctcagc
      241 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      301 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      361 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      421 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      481 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
      541 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      601 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      661 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      721 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      781 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
      841 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      901 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      961 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1021 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1081 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1141 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1201 ctaagaaacc attattatca taaaaggcca tccgtcagga tggccttcta tgaatgcctt
     1261 ggctgcgatt cattctttat atgaataaaa ttgttgtcag tttcacgtct tgtcctgcca
     1321 tatcgcgaaa tttctgcgca aaagcacaaa aaatttttgc atctccccct tgatgacgtg
     1381 ggttacgacc ccatttagta gtcaaccgca gtgagtgagt ctgacgaggc cctttcgtct
     1441 tcaagaattc tcatgtttga cagcttatca tcgataagct ttaatgcggt agtttatcac
     1501 agttaaattg ctaacgcagt ccggggaatt caggcttgga ggaacaatat gaaaggatcc
     1561 aaaggtgaag aattattcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     1621 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     1681 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     1741 actttcgcgt atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac
     1801 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     1861 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     1921 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     1981 tacaactata actcacacaa agtatacatc atggcagaca aacaaaagga tggaatcaaa
     2041 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     2101 caacgaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2161 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2221 tttgtaacag ctgctgggat tacacatggt atggatgaat tgtacaaata accgccagga
     2281 ggaacaatat ggtgagcaag ggcaaaaaga ggcagtgatc aaggagttca tgcggttcaa
     2341 ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2401 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2461 cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct tcatcaagca
     2521 ccccgccgac atccccgact actataagca gtccttcccc gagggcttca agtgggagcg
     2581 cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga
     2641 cggcaccctg atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt
     2701 aatgcagaag aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg
     2761 cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc
     2821 ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt
     2881 cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga
     2941 acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt gaccgcctaa
     3001 tgagcgggct ttaatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag
     3061 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccacc accactaatt
     3121 gattaatacc taggctgcta aacaaagccc atggatcgat agctggtcga caggactctg
     3181 gggttcgaga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     3241 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     3301 actagggaca gtaagacggg taagcctgtt gatgataccg ctgccttact gggtgcatta
     3361 gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa aatcagaggg
     3421 caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg ccataaaacg
     3481 ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg catgaatcca
     3541 taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga gcgcagcgaa
     3601 ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca aaaggaatat
     3661 tcagcgattt gcccgagctt gcgagggtgc tacttaagcc tttagggttt taaggtctgt
     3721 tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga actgactaaa
     3781 gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat tctttcttta
     3841 ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt ctagcggaat
     3901 ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag aatttacaga
     3961 tacccacaac tcaaaggaaa aggactagta attatcattg actagcccat ctcaattggt
     4021 atagtgatta aaatcaccta gaccaattga gatgtatgtc tgaattagtt gttttcaaag
     4081 caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag ctaattttat
     4141 gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa cggacggtat
     4201 cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat gcttatggtg
     4261 tattagctaa agcaaccaga gagctgatga cgagaactgt ggaaatcagg aatcctttgg
     4321 ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc gaaaaattag
     4381 aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa ttcataaaat
     4441 ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt tatgagtggt
     4501 tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt agccttgatg
     4561 aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg cttaaccaat
     4621 gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa ttggtggttg
     4681 ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat agacaaatgg
     4741 atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa ataccaacaa
     4801 ccattacatc agattcctac ctacataacg gactaagaaa aacactacac gatgctttaa
     4861 ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg caaagtaagt
     4921 atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca ctagagaaca
     4981 tactggctaa atacggaagg atctgaggtt cttatggctc ttgtatctat cagtgaagca
     5041 tcaagactaa caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg
     5101 tccatatgca cagatgaaaa cggtgtaaaa aagatagata catcagagct tttacgagtt
     5161 tttggtgcat tcaaagctgt tcaccatgaa cagatcgaca atgtaacaga tgaacagcat
     5221 gtaacaccta atagaacagg tgaaaccagt aaaacaaagc aactagaaca tgaaattgaa
     5281 cacctgagac aacttgttac agctcaacag tcacacatag acagcctgaa acaggcgatg
     5341 ctgcttatcg aatcaaagct gccgacaaca cgggagccag tgacgcctcc cgtggggaaa
     5401 aaatcatggc aattctggaa gaaatagcgc tttcagccgg caaaccggct gaagccggat
     5461 ctgcgattct gataacaaac tagcaacacc agaacagccc gtttgcgggc agcaaaaccc
     5521 gtaccctagg tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     5581 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg a
//
