LOCUS       sequence-358101-        4422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression construct encoding 3xFLAG and mCherry from separate 
            promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4422)
  AUTHORS   Lee S, Park D, Lim C, Kim JI, Min KT
  TITLE     mtIF3 is locally translated in axons and regulates mitochondrial 
            translation for axonal growth.
  JOURNAL   BMC Biol. 2022 Jan 7;20(1):12. doi: 10.1186/s12915-021-01215-w.
  PUBMED    34996455
REFERENCE   2  (bases 1 to 4422)
  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: "BMC Biol.";
            date: "2022-01-7"; volume: "20(1):12. doi"; pages: " 
            10.1186/s12915-021-01215-w"
COMMENT     Sequence Label: psiCheck2-SV40p-3xFLAG-HSVTKp-mCherry
FEATURES             Location/Qualifiers
     source          1..4422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             693..758
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    777..825
                     /note="synthetic polyadenylation signal"
     promoter        834..1585
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             1627..2337
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1772..1790)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2040..2059
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2377..2498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2388..2407)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2442..2461
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2631..2735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2736..2804,2805..3596)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2954..2973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3767..4355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4256..4275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagca tggactataa ggaccacgac ggagactaca
      721 aggatcatga tattgattac aaagacgatg acgataagta agcggccgct ggccgcaata
      781 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt
      841 cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg
      901 gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg
      961 gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg
     1021 tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt
     1081 gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc
     1141 gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga
     1201 ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga
     1261 gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag
     1321 gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt
     1381 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
     1441 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
     1501 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     1561 agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc
     1621 gggcccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1681 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1741 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1801 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1861 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1921 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1981 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2041 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2101 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2161 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2221 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2281 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtagtct
     2341 agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     2401 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2461 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2521 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2581 aatgtggtaa aatcgataag gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2641 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2701 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2761 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2821 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2881 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2941 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3001 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3061 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3121 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3181 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3241 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3301 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3361 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3421 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3481 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3541 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3601 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3661 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3721 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3781 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3841 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3901 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3961 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4021 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4081 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4141 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4201 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4261 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4321 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4381 ggttcctggc cttttgctgg ccttttgctc acatggctcg ac
//
