LOCUS       sequence-180166-        4951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses shRNA against TOM40..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4951)
  AUTHORS   Yano H, Baranov SV, Baranova OV, Kim J, Pan Y, Yablonska S, Carlisle
            DL, Ferrante RJ, Kim AH, Friedlander RM
  TITLE     Inhibition of mitochondrial protein import by mutant huntingtin.
  JOURNAL   Nat Neurosci. 2014 Jun;17(6):822-31. doi: 10.1038/nn.3721. Epub 2014
            May 18.
  PUBMED    24836077
REFERENCE   2  (bases 1 to 4951)
  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"; doi: "10.1038/nn.3721"; 
            journalName: "Nat Neurosci"; date: "2014-06"; volume: "17"; issue: 
            "6"; pages: "822-31"
COMMENT     Sequence Label: pBSU6-shTom40.3/CMV-eGFP
FEATURES             Location/Qualifiers
     source          1..4951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     280..297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    414..435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(450..455,456..473,474..480)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    488..504
                     /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)."
     primer_bind     493..515
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     512..528
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     512..528
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     547..567
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        549..567
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     604..620
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    653..734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     678..697
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(857..1570,1571..1573,1574..1576))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(860..1570,1571..1573,1574..1576))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(902..923)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1251..1270
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1510..1531
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1573..1582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1584..1664
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(1687..1890)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1716..1736)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1891..2194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(2342..2655)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(2403..2423)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(2652..2668)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2685..2704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2686..2704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2714..2731)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2714..2730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2723..2745)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2872..3327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2959..2978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3169..3190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3353..3457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3458..3526,3527..4318)
                     /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(3676..3695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4489..4951,1..126)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
       61 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      301 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      361 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      421 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      481 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      541 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctccaccg cggtggcggc
      601 cgctctagaa ctagtggatc ccccggaagt ttggacaaac cacaactaga atgcagtgaa
      661 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      721 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      781 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
      841 tagagtcgcg gccgctttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
      901 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
      961 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     1021 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     1081 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     1141 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1201 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1261 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1321 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     1381 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     1441 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1501 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1561 cgcccttgct caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca
     1621 gaattcgaag cttgagctcg agatctgagt ccggtagcgc tagcggatct gacggttcac
     1681 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     1741 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     1801 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     1861 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     1921 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     1981 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2041 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2101 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2161 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     2221 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata ccctgcagga
     2281 attcaaaaag gcactgtcat gtctctagct aagcttgaat gctagagaca tgacagtgcc
     2341 aaacaaggct tttctccaag ggatatttat agtctcaaaa cacacaatta ctttacagtt
     2401 agggtgagtt tccttttgtg ctgtttttta aaataataat ttagtatttg tatctcttat
     2461 agaaatccaa gcctatcatg taaaatgtag ctagtattaa aaagaacaga ttatctgtct
     2521 tttatcgcac attaagcctc tatagttact aggaaatatt atatgcaaat taaccggggc
     2581 aggggagtag ccgagcttct cccacaagtc tgtgcgaggg ggccggcgcg ggcctagaga
     2641 tggcggcgtc ggatccacta gttctagagc gggtacccaa ttcgccctat agtgagtcgt
     2701 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2761 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2821 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2941 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3061 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3181 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3241 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3301 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3361 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3421 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3481 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3541 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3601 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3661 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3721 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3781 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3841 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3901 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3961 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4021 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4081 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4141 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4201 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4261 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4321 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4381 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4621 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4921 caggtatccg gtaagcggca gggtcggaac a
//
