LOCUS       sequence-358105-        5253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression construct encoding shRNA-resistant mtIF3 and mCherry
            from separate promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5253)
  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 5253)
  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-mtIF3(SR)-3xFLAG-HSVTKp-mCherry
FEATURES             Location/Qualifiers
     source          1..5253
                     /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             1524..1589
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1608..1656
                     /note="synthetic polyadenylation signal"
     promoter        1665..2416
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2458..3168
                     /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(2603..2621)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2871..2890
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3208..3329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3219..3238)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3273..3292
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3462..3566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3567..3635,3636..4427)
                     /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(3785..3804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4598..5186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5087..5106
                     /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 tggctgttct tctgatgagg ttgatgctac
      721 aaactacaaa gcttgaccat aacttaattg gaagatgttt gcaaagacac gctgtgaagc
      781 cagatccagc acagctgtcc ctgtctgcgt ccaccccaaa gctgttgtac ctgacttctg
      841 caaaaggttt tagtactgct ggagacccac agggtgagag gaaacagaaa agaagggatg
      901 ctttctctaa caccggccgg aagattagtg agcggattat ccgcgtcctg gatgagaagg
      961 gcatggacct ggggatgatg catcgagcag atgtgatcag actcatgaac aagcaggacc
     1021 tgaggctggt gcagaggaac accagctcag agccaccgga gtaccagctt atgaccggag
     1081 agcagatcca ccaggagcgg ctaaagctcc gggaacagga aaaggccaaa cccaaaactg
     1141 gaccaactat gacgaaagag ctcgttttct cttcaaatat tggacagcac gatttagaca
     1201 ccaagagtaa acagatccag cagtggatag agaagaaata tcatgtacag gtgaccatta
     1261 agaggaggaa agatgcggag cagtcagagg aggaaacgga ggagatattt aaccagattc
     1321 tccagactat gcctgacata gcaaccttct caagcagacc aaaagccatc agaggaggga
     1381 ctgcttccat gtgtgttttt cgtcacttga gcaagaaaga agaaaaggcg tacagagaat
     1441 cccaggagag ccagagaaga gacactttga gcaaagacga cgacggaaac agcaaagagt
     1501 cggatgttgt gtgtcagacc ggtgactata aggaccacga cggagactac aaggatcatg
     1561 atattgatta caaagacgat gacgataagt aagcggccgc tggccgcaat aaaatatctt
     1621 tattttcatt acatctgtgt gttggttttt tgtgtgagga tctaaatgag tcttcggacc
     1681 tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac gcggggggag ggggaaggaa
     1741 cgaaacactc tcattcggag gcggctcggg gtttggtctt ggtggccacg ggcacgcaga
     1801 agagcgccgc gatcctctta agcacccccc cgccctccgt ggaggcgggg gtttggtcgg
     1861 cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg cgccccagag tgtgaccttt
     1921 tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg gcgacgggct cgctgggtcc
     1981 taggctccat ggggaccgta tacgtggaca ggctctggag catccgcacg actgcggtga
     2041 tattaccgga gaccttctgc gggacgagcc gggtcacgcg gctgacgcgg agcgtccgtt
     2101 gggcgacaaa caccaggacg gggcacaggt acactatctt gtcacccgga ggcgcgaggg
     2161 actgcaggag cttcagggag tggcgcagct gcttcatccc cgtggcccgt tgctcgcgtt
     2221 tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa
     2281 ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg
     2341 gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct
     2401 gcagcgaccc gcttaaaagc ttggcattcc ggtactgttg gtaaagccac cgggcccatg
     2461 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2521 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2581 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2641 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2701 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2761 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2821 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2881 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2941 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     3001 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     3061 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     3121 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtagtc tagagtcggg
     3181 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3241 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3301 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3361 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3421 aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3481 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3541 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3601 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3661 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3721 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3781 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3841 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3901 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3961 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4021 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4081 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4141 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4201 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4261 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4321 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4381 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4441 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4501 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4561 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4621 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4681 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4741 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4801 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4861 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4921 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4981 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5041 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5101 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5161 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5221 ccttttgctg gccttttgct cacatggctc gac
//
