LOCUS       sequence-345057-        8260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitochondria targeted dsRED expressed off of EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8260)
  TITLE     Fluorescent plasmids for live cell imaging in primary neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8260)
  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            "Unpublished"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: EF1a-mito-dsRED2
FEATURES             Location/Qualifiers
     source          1..8260
                     /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)"
     promoter        2221..2432
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2433..3371
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3328..3348
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(3382..3398)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3382..3398)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3419..3493
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      3521..3530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3527..4204
                     /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(3707..3727)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4119..4142
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    4229..4817
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4282..4302)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4700..4711)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5024..5657
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5785..5801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5785..5801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5798..5820)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5809..5825
                     /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(5833..5839,5840..5857,5858..5863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5878..5899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6016..6033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6187..6775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6267..6286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6946..7737,7738..7806))
                     /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     7569..7588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7807..7911)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7959..8093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7996..8015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8050..8069
                     /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 cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt
     2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3361 ttccatttca ggtgtcgtga ggatccacta gttctagagc ggccgcagat ccgctagcat
     3421 gtccgtcctg acgccgctgc tgctgcgggg cttgacaggc tcggcccggc ggctcccagt
     3481 gccgcgcgcc aagatccatt cgttggggga tccaccggtc gccaccatgg cctcctccga
     3541 gaacgtcatc accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca
     3601 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa
     3661 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt
     3721 ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct
     3781 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc
     3841 gaccgtgacc caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat
     3901 cggcgtgaac ttcccctccg acggccccgt gatgcagaag aagaccatgg gctgggaggc
     3961 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct
     4021 gaagctgaag gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa
     4081 gcccgtgcag ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa
     4141 cgaggactac accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct
     4201 gtagtgtaca agtgaacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag
     4261 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4321 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4381 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4441 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4501 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4561 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4621 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4681 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4741 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4801 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca
     4861 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     4921 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     4981 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac
     5041 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     5101 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     5161 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     5221 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     5281 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     5341 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     5401 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     5461 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5521 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5581 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5641 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     5701 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc
     5761 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5821 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5881 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5941 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6001 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7921 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata
     7981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     8041 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     8101 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     8161 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
     8221 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
