LOCUS       sequence-417987-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mistranslation GFP and mScarlet-I positive control with Trp 
            insertion.
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 6Trp7Ser
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(191..982,983..1051))
                     /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     814..833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1052..1156)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1208..1235)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1443..1448,1449..1465,1466..1471)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1571..1599)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2194..2214
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2275..2976
                     /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="MVSKGWSEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFED
                     GGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGD
                     IKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEG
                     RHSTGGMDELYK"
     primer_bind     complement(2414..2432)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2682..2701
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3079..3102
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3187..3281
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3773..3995
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4043..4993
                     /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(5531..5617)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcatgcccaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
       61 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      121 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      181 cggcgtagag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      481 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
      541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      781 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
      841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1141 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     1201 ttatcataaa aggccatccg tcaggatggc cttctatgaa tgccttggct gcgattcatt
     1261 ctttatatga ataaaattgt tgtcagtttc acgtcttgtc ctgccatatc gcgaaatttc
     1321 tgcgcaaaag cacaaaaaat ttttgcatct cccccttgat gacgtgggtt acgaccccat
     1381 ttagtagtca accgcagtga gtgagtctga cgaggccctt tcgtcttcaa gaattctcat
     1441 gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt aaattgctaa
     1501 cgcagtccgg ggaattcagg cttggaggaa caatatgaaa ggatccaaag gtgaagaatt
     1561 attcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     1621 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     1681 ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt tcgcgtatgg
     1741 tcttcaatgc tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc
     1801 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
     1861 gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     1921 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     1981 acacaaagta tacatcatgg cagacaaaca aaaggatgga atcaaagtta acttcaaaat
     2041 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac gaaatactcc
     2101 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     2161 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     2221 tgggattaca catggtatgg atgaattgta caaataaccg ccaggaggaa caatatggtg
     2281 agcaagggct ggagtgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
     2341 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2401 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
     2461 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
     2521 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2581 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     2641 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
     2701 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     2761 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
     2821 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     2881 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
     2941 cgccactcca ccggcggcat ggacgagctg tacaagtgac cgcctaatga gcgggcttta
     3001 atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc cgtcgacaag
     3061 cttgcggccg cactcgagca ccaccaccac caccaccacc actaattgat taatacctag
     3121 gctgctaaac aaagcccatg gatcgatagc tggtcgacag gactctgggg ttcgagagct
     3181 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     3241 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact agggacagta
     3301 agacgggtaa gcctgttgat gataccgctg ccttactggg tgcattagcc agtctgaatg
     3361 acctgtcacg ggataatccg aagtggtcag actggaaaat cagagggcag gaactgctga
     3421 acagcaaaaa gtcagatagc accacatagc agacccgcca taaaacgccc tgagaagccc
     3481 gtgacgggct tttcttgtat tatgggtagt ttccttgcat gaatccataa aaggcgcctg
     3541 tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga
     3601 aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgatttgcc
     3661 cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc
     3721 aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata
     3781 cacagggctg ggatctattc tttttatctt tttttattct ttctttattc tataaattat
     3841 aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct
     3901 agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca
     3961 aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata gtgattaaaa
     4021 tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc
     4081 gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact
     4141 actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa
     4201 ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc
     4261 aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggctttga
     4321 gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag
     4381 tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca
     4441 tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact
     4501 aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat
     4561 gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc
     4621 aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg
     4681 cccgactgat acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga
     4741 acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga
     4801 ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg caaaaattca
     4861 gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg atctcaatgg
     4921 ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata
     4981 cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa
     5041 acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc atatgcacag
     5101 atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt ggtgcattca
     5161 aagctgttca ccatgaacag atcgacaatg taacagatga acagcatgta acacctaata
     5221 gaacaggtga aaccagtaaa acaaagcaac tagaacatga aattgaacac ctgagacaac
     5281 ttgttacagc tcaacagtca cacatagaca gcctgaaaca ggcgatgctg cttatcgaat
     5341 caaagctgcc gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa tcatggcaat
     5401 tctggaagaa atagcgcttt cagccggcaa accggctgaa gccggatctg cgattctgat
     5461 aacaaactag caacaccaga acagcccgtt tgcgggcagc aaaacccgta ccctaggtct
     5521 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     5581 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagagc tt
//
