LOCUS       sequence-419242-        4875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP positive, mScarlet-I negative control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4875)
  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 4875)
  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(Delta-mScarlet)
FEATURES             Location/Qualifiers
     source          1..4875
                     /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
     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             2323..2346
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      2431..2525
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3017..3239
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3287..4237
                     /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(4775..4861)
                     /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 tgacggatgg 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 gagcaagggg catggacgag ctgtacaagt aaaagcctcg agcaccacca ccaccaccac
     2341 caccactaat tgattaatac ctaggctgct aaacaaagcc catggatcga tagctggtcg
     2401 acaggactct ggggttcgag agctcgcttg gactcctgtt gatagatcca gtaatgacct
     2461 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     2521 agaatccaag cactagggac agtaagacgg gtaagcctgt tgatgatacc gctgccttac
     2581 tgggtgcatt agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga
     2641 aaatcagagg gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc
     2701 gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt
     2761 gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg
     2821 agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac
     2881 aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt
     2941 ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg
     3001 aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta
     3061 ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg
     3121 tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca
     3181 gaatttacag atacccacaa ctcaaaggaa aaggactagt aattatcatt gactagccca
     3241 tctcaattgg tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt
     3301 tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa
     3361 gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga
     3421 acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa
     3481 tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag
     3541 gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag
     3601 cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa
     3661 attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat
     3721 ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat
     3781 tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag
     3841 gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa
     3901 attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga
     3961 tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa
     4021 aataccaaca accattacat cagattccta cctacataac ggactaagaa aaacactaca
     4081 cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat
     4141 gcaaagtaag tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac
     4201 actagagaac atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta
     4261 tcagtgaagc atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa
     4321 agggaaaact gtccatatgc acagatgaaa acggtgtaaa aaagatagat acatcagagc
     4381 ttttacgagt ttttggtgca ttcaaagctg ttcaccatga acagatcgac aatgtaacag
     4441 atgaacagca tgtaacacct aatagaacag gtgaaaccag taaaacaaag caactagaac
     4501 atgaaattga acacctgaga caacttgtta cagctcaaca gtcacacata gacagcctga
     4561 aacaggcgat gctgcttatc gaatcaaagc tgccgacaac acgggagcca gtgacgcctc
     4621 ccgtggggaa aaaatcatgg caattctgga agaaatagcg ctttcagccg gcaaaccggc
     4681 tgaagccgga tctgcgattc tgataacaaa ctagcaacac cagaacagcc cgtttgcggg
     4741 cagcaaaacc cgtaccctag gtctagggcg gcggatttgt cctactcagg agagcgttca
     4801 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     4861 gatgcctcta gagct
//
