LOCUS       sequence-416760-        5626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mistranslation GFP and mScarlet-I positive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5626)
  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 5626)
  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
FEATURES             Location/Qualifiers
     source          1..5626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(196..987,988..1056))
                     /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     819..838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1057..1161)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1213..1240)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1448..1453,1454..1470,1471..1476)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1576..1604)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2199..2219
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2280..2975
                     /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="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2413..2431)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2681..2700
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3078..3101
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3186..3280
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3772..3994
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4042..4992
                     /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(5530..5616)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcttgcatg cccaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
       61 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
      121 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      181 gcgtccggcg tagagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      481 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
      541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      781 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
      841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1201 cattattatc ataaaaggcc atccgtcagg atggccttct atgaatgcct tggctgcgat
     1261 tcattcttta tatgaataaa attgttgtca gtttcacgtc ttgtcctgcc atatcgcgaa
     1321 atttctgcgc aaaagcacaa aaaatttttg catctccccc ttgatgacgt gggttacgac
     1381 cccatttagt agtcaaccgc agtgagtgag tctgacgagg ccctttcgtc ttcaagaatt
     1441 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
     1501 gctaacgcag tccggggaat tcaggcttgg aggaacaata tgaaaggatc caaaggtgaa
     1561 gaattattca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     1621 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     1681 tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcgcg
     1741 tatggtcttc aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag
     1801 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
     1861 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     1921 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     1981 aactcacaca aagtatacat catggcagac aaacaaaagg atggaatcaa agttaacttc
     2041 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacgaaat
     2101 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     2161 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     2221 gctgctggga ttacacatgg tatggatgaa ttgtacaaat aaccgccagg aggaacaata
     2281 tggtgagcaa gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg
     2341 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2401 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca
     2461 tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc
     2521 ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2581 aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct
     2641 acaaggtgaa gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga
     2701 caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg
     2761 acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca
     2821 cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg
     2881 acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc
     2941 gccactccac cggcggcatg gacgagctgt acaagtgacc gcctaatgag cgggctttaa
     3001 tcactagtga attcgcggcc gcctgcaggt cgaccatatg ggagagctcc gtcgacaagc
     3061 ttgcggccgc actcgagcac caccaccacc accaccacca ctaattgatt aatacctagg
     3121 ctgctaaaca aagcccatgg atcgatagct ggtcgacagg actctggggt tcgagagctc
     3181 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     3241 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta gggacagtaa
     3301 gacgggtaag cctgttgatg ataccgctgc cttactgggt gcattagcca gtctgaatga
     3361 cctgtcacgg gataatccga agtggtcaga ctggaaaatc agagggcagg aactgctgaa
     3421 cagcaaaaag tcagatagca ccacatagca gacccgccat aaaacgccct gagaagcccg
     3481 tgacgggctt ttcttgtatt atgggtagtt tccttgcatg aatccataaa aggcgcctgt
     3541 agtgccattt acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa
     3601 aaagacagcg actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgatttgccc
     3661 gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca
     3721 aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac
     3781 acagggctgg gatctattct ttttatcttt ttttattctt tctttattct ataaattata
     3841 accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta
     3901 gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa
     3961 aggaaaagga ctagtaatta tcattgacta gcccatctca attggtatag tgattaaaat
     4021 cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg
     4081 attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta
     4141 ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac
     4201 caatacgctc agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca
     4261 accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttgag
     4321 attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt
     4381 gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat
     4441 gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta
     4501 acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg
     4561 ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca
     4621 ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc
     4681 ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa
     4741 cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat
     4801 tcctacctac ataacggact aagaaaaaca ctacacgatg ctttaactgc aaaaattcag
     4861 ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt
     4921 tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac
     4981 ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa
     5041 caaaagtaga acaactgttc accgttacat atcaaaggga aaactgtcca tatgcacaga
     5101 tgaaaacggt gtaaaaaaga tagatacatc agagctttta cgagtttttg gtgcattcaa
     5161 agctgttcac catgaacaga tcgacaatgt aacagatgaa cagcatgtaa cacctaatag
     5221 aacaggtgaa accagtaaaa caaagcaact agaacatgaa attgaacacc tgagacaact
     5281 tgttacagct caacagtcac acatagacag cctgaaacag gcgatgctgc ttatcgaatc
     5341 aaagctgccg acaacacggg agccagtgac gcctcccgtg gggaaaaaat catggcaatt
     5401 ctggaagaaa tagcgctttc agccggcaaa ccggctgaag ccggatctgc gattctgata
     5461 acaaactagc aacaccagaa cagcccgttt gcgggcagca aaacccgtac cctaggtcta
     5521 gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     5581 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctctag
//
