LOCUS       sequence-419244-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  frameshift mistranslation, A6T.
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 A6T
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(202..993,994..1062))
                     /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     825..844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1063..1167)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1219..1246)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1454..1459,1460..1476,1477..1482)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1582..1610)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2205..2225
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     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 ctctagagct tgcatgccca aggagatggc gcccaacagt cccccggcca cggggcctgc
       61 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
      121 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      181 cacgatgcgt ccggcgtaga gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      241 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      301 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      361 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      421 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      481 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
      541 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      601 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      661 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      721 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      781 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
      841 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      901 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      961 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1021 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1081 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1141 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     1201 agaaaccatt attatcataa aaggccatcc gtcaggatgg ccttctatga atgccttggc
     1261 tgcgattcat tctttatatg aataaaattg ttgtcagttt cacgtcttgt cctgccatat
     1321 cgcgaaattt ctgcgcaaaa gcacaaaaaa tttttgcatc tcccccttga tgacgtgggt
     1381 tacgacccca tttagtagtc aaccgcagtg agtgagtctg acgaggccct ttcgtcttca
     1441 agaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt
     1501 taaattgcta acgcagtccg gggaattcag gcttggagga acaatatgaa aggatccaaa
     1561 ggtgaagaat tattcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     1621 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     1681 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     1741 ttcgcgtatg gtcttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt
     1801 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     1861 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     1921 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     1981 aactataact cacacaaagt atacatcatg gcagacaaac aaaaggatgg aatcaaagtt
     2041 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     2101 cgaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     2161 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     2221 gtaacagctg ctgggattac acatggtatg gatgaattgt acaaataacc gccaggagga
     2281 acaatatggt gagcaagggc aaaaaatgag 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 tgc
//
