LOCUS       sequence-417986-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nonsense mistranslation, TGA stop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5632)
  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 5632)
  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 TGA
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(192..983,984..1052))
                     /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     815..834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1053..1157)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1209..1236)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1444..1449,1450..1466,1467..1472)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1572..1600)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2195..2215
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2276..2977
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKG*SEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFED
                     GGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGD
                     IKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEG
                     RHSTGGMDELYK"
     primer_bind     complement(2415..2433)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2683..2702
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3080..3103
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3188..3282
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3774..3996
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4044..4994
                     /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(5532..5618)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcatgccca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
       61 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
      121 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      181 ccggcgtaga gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
      241 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
      301 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
      361 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
      421 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      481 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
      541 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
      601 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
      661 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      721 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      781 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
      841 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      901 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      961 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     1021 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     1081 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1141 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     1201 attatcataa aaggccatcc gtcaggatgg ccttctatga atgccttggc tgcgattcat
     1261 tctttatatg aataaaattg ttgtcagttt cacgtcttgt cctgccatat cgcgaaattt
     1321 ctgcgcaaaa gcacaaaaaa tttttgcatc tcccccttga tgacgtgggt tacgacccca
     1381 tttagtagtc aaccgcagtg agtgagtctg acgaggccct ttcgtcttca agaattctca
     1441 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
     1501 acgcagtccg gggaattcag gcttggagga acaatatgaa aggatccaaa ggtgaagaat
     1561 tattcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     1621 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
     1681 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcgcgtatg
     1741 gtcttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt ttcaagagtg
     1801 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
     1861 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     1921 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     1981 cacacaaagt atacatcatg gcagacaaac aaaaggatgg aatcaaagtt aacttcaaaa
     2041 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa cgaaatactc
     2101 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     2161 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     2221 ctgggattac acatggtatg gatgaattgt acaaataacc gccaggagga acaatatggt
     2281 gagcaagggc tgaagtgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga
     2341 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2401 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga
     2461 catcctgtcc cctcagttca tgtacggctc cagggccttc atcaagcacc ccgccgacat
     2521 ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2581 cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat
     2641 ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa
     2701 gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg
     2761 cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg acttcaagac
     2821 cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt
     2881 ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg
     2941 ccgccactcc accggcggca tggacgagct gtacaagtga ccgcctaatg agcgggcttt
     3001 aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct ccgtcgacaa
     3061 gcttgcggcc gcactcgagc accaccacca ccaccaccac cactaattga ttaataccta
     3121 ggctgctaaa caaagcccat ggatcgatag ctggtcgaca ggactctggg gttcgagagc
     3181 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     3241 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac tagggacagt
     3301 aagacgggta agcctgttga tgataccgct gccttactgg gtgcattagc cagtctgaat
     3361 gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca ggaactgctg
     3421 aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc ctgagaagcc
     3481 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct
     3541 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg
     3601 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc
     3661 ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt tgtagaggag
     3721 caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt agtgagttat
     3781 acacagggct gggatctatt ctttttatct ttttttattc tttctttatt ctataaatta
     3841 taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt acagagggtc
     3901 tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata cccacaactc
     3961 aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat agtgattaaa
     4021 atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca aatgaactag
     4081 cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc tgtgtggcac
     4141 tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg ttcacttata
     4201 accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta ttagctaaag
     4261 caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt aaaggctttg
     4321 agattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa ttagttttta
     4381 gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat aatctggaac
     4441 atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta ttaaaagaac
     4501 taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa tttaagttca
     4561 tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg gttttgaaac
     4621 caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat aagcgaggcc
     4681 gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat ctcgtaaccg
     4741 aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc attacatcag
     4801 attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact gcaaaaattc
     4861 agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat gatctcaatg
     4921 gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata ctggctaaat
     4981 acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc aagactaaca
     5041 aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc catatgcaca
     5101 gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt tggtgcattc
     5161 aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt aacacctaat
     5221 agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca cctgagacaa
     5281 cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct gcttatcgaa
     5341 tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa atcatggcaa
     5401 ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct gcgattctga
     5461 taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt accctaggtc
     5521 tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     5581 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagag ct
//
