LOCUS       sequence-345054-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondria targeted dsRED2 expressed off of CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  TITLE     Fluorescent plasmids for live cell imaging in primary neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7872)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CAG-mito-dsRED2
FEATURES             Location/Qualifiers
     source          1..7872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     intron          2830..2926
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             3031..3105
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      3133..3142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3139..3816
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(3319..3339)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3731..3754
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3841..4429
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3894..3914)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4312..4323)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4636..5269
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5397..5413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5397..5413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5410..5432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5421..5437
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5445..5451,5452..5469,5470..5475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5490..5511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5628..5645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5799..6387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5879..5898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6558..7349,7350..7418))
                     /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     7181..7200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7419..7523)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7571..7705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7608..7627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7662..7681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgattaatta aggtacctct ggtcgttaca taacttacgg
     2221 taaatggccc gcctggctga ccgcccaacg accccgccca ttgacgtcaa taatgacgta
     2281 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     2341 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2401 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2461 tcctacttgg cagtacatct actcgaggcc acgttctgct tcactctccc catctccccc
     2521 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2581 gcgggggggg gggggggggg ggcgcgcgcs rggsggggsg gggsggggsg rggggcgggg
     2641 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     2701 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     2761 agcgggatca gccaccgcgg tggcggccct agagtcgatc gaggaactga aaaaccagaa
     2821 agttaactgg taagtttagt ctttttgtct tttatttcag gtccccgatc cggtggtggt
     2881 gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt
     2941 gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgatccgaat tcctcgagac
     3001 tagttctaga gcggccgcag atccgctagc atgtccgtcc tgacgccgct gctgctgcgg
     3061 ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca ttcgttgggg
     3121 gatccaccgg tcgccaccat ggcctcctcc gagaacgtca tcaccgagtt catgcgcttc
     3181 aaggtgcgca tggagggcac cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     3241 ggccgcccct acgagggcca caacaccgtg aagctgaagg tgaccaaggg cggccccctg
     3301 cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag
     3361 caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag
     3421 cgcgtgatga acttcgagga cggcggcgtg gcgaccgtga cccaggactc ctccctgcag
     3481 gacggctgct tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc
     3541 gtgatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     3601 ggcgtgctga agggcgagac ccacaaggcc ctgaagctga aggacggcgg ccactacctg
     3661 gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac
     3721 gtggacgcca agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac
     3781 gagcgcaccg agggccgcca ccacctgttc ctgtagtgta caagtgaacg cgtctggaac
     3841 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3901 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3961 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4021 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4081 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4141 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4201 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4261 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4321 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4381 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
     4441 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     4501 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     4561 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4621 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     4681 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
     4741 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
     4801 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
     4861 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
     4921 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
     4981 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
     5041 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
     5101 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5161 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5221 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     5281 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     5341 ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     5401 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5461 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5521 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6541 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6601 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6661 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6721 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6781 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6841 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6901 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6961 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7021 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7081 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7141 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7201 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7261 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7321 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7381 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7441 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7501 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7561 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7681 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     7741 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     7801 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     7861 aaacttagta gt
//
