LOCUS       sequence-417835-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  frameshift mistranslation, T7.
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 T7
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..991
                     /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(1529..1615)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1743..1762
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(1821..2612,2613..2681))
                     /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     2444..2463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2682..2786)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2838..2865)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(3073..3078,3079..3095,3096..3101)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3201..3229)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3824..3844
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4045..4063)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4313..4332
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4710..4733
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      4818..4912
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5404..5626
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttggtatagt gattaaaatc acctagacca attgagatgt atgtctgaat tagttgtttt
       61 caaagcaaat gaactagcga ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat
      121 tttatgctgt gtggcactac tcaaccccac gattgaaaac cctacaagga aagaacggac
      181 ggtatcgttc acttataacc aatacgctca gatgatgaac atcagtaggg aaaatgctta
      241 tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc
      301 tttggttaaa ggctttgaga ttttccagtg gacaaactat gccaagttct caagcgaaaa
      361 attagaatta gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat
      421 aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga
      481 gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct
      541 tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa
      601 ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt
      661 ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca
      721 aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc
      781 aacaaccatt acatcagatt cctacctaca taacggacta agaaaaacac tacacgatgc
      841 tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag
      901 taagtatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga
      961 gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg
     1021 aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata tcaaagggaa
     1081 aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac
     1141 gagtttttgg tgcattcaaa gctgttcacc atgaacagat cgacaatgta acagatgaac
     1201 agcatgtaac acctaataga acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa
     1261 ttgaacacct gagacaactt gttacagctc aacagtcaca catagacagc ctgaaacagg
     1321 cgatgctgct tatcgaatca aagctgccga caacacggga gccagtgacg cctcccgtgg
     1381 ggaaaaaatc atggcaattc tggaagaaat agcgctttca gccggcaaac cggctgaagc
     1441 cggatctgcg attctgataa caaactagca acaccagaac agcccgtttg cgggcagcaa
     1501 aacccgtacc ctaggtctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca
     1561 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     1621 tctagagctt gcatgcccaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     1681 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     1741 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     1801 acgatgcgtc cggcgtagag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2101 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     2161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     2461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2821 gaaaccatta ttatcataaa aggccatccg tcaggatggc cttctatgaa tgccttggct
     2881 gcgattcatt ctttatatga ataaaattgt tgtcagtttc acgtcttgtc ctgccatatc
     2941 gcgaaatttc tgcgcaaaag cacaaaaaat ttttgcatct cccccttgat gacgtgggtt
     3001 acgaccccat ttagtagtca accgcagtga gtgagtctga cgaggccctt tcgtcttcaa
     3061 gaattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
     3121 aaattgctaa cgcagtccgg ggaattcagg cttggaggaa caatatgaaa ggatccaaag
     3181 gtgaagaatt attcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
     3241 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
     3301 ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt
     3361 tcgcgtatgg tcttcaatgc tttgcgagat acccagatca tatgaaacag catgactttt
     3421 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
     3481 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
     3541 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
     3601 actataactc acacaaagta tacatcatgg cagacaaaca aaaggatgga atcaaagtta
     3661 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
     3721 gaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
     3781 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
     3841 taacagctgc tgggattaca catggtatgg atgaattgta caaataaccg ccaggaggaa
     3901 caatatggtg agcaagggct ttttttgagg cagtgatcaa ggagttcatg cggttcaagg
     3961 tgcacatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     4021 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     4081 tctcctggga catcctgtcc cctcagttca tgtacggctc cagggccttc atcaagcacc
     4141 ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg
     4201 tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg
     4261 gcaccctgat ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa
     4321 tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg
     4381 tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg
     4441 acttcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg
     4501 accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac
     4561 gctccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga ccgcctaatg
     4621 agcgggcttt aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct
     4681 ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccaccac cactaattga
     4741 ttaataccta ggctgctaaa caaagcccat ggatcgatag ctggtcgaca ggactctggg
     4801 gttcgagagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
     4861 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac
     4921 tagggacagt aagacgggta agcctgttga tgataccgct gccttactgg gtgcattagc
     4981 cagtctgaat gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca
     5041 ggaactgctg aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc
     5101 ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata
     5161 aaaggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact
     5221 gaatgtcacg aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc
     5281 agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt
     5341 tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt
     5401 agtgagttat acacagggct gggatctatt ctttttatct ttttttattc tttctttatt
     5461 ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt
     5521 acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata
     5581 cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct caa
//
