LOCUS       sequence-201359-        3814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3814)
  AUTHORS   Maynard CJ, Boettcher C, Ortega Z, Smith R, Florea BI, 
            Diaz-Hernandez M, Brundin P, Overkleeft HS, Li JY, Lucas JJ, Dantuma
            NP
  TITLE     Accumulation of ubiquitin conjugates in a polyglutamine disease 
            model occurs without global ubiquitin/proteasome system impairment.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Aug 18. 106(33):13986-91.
  PUBMED    19666572
REFERENCE   2  (bases 1 to 3814)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Aug 18. 106(33):13986-91."
COMMENT     Sequence Label: pTreTight-Htt94Q-CFP
FEATURES             Location/Qualifiers
     source          1..3814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    109..190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(146..165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(201..218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(372..960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(452..471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1131..1922,1923..1991))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1754..1773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1992..2096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2164..2182
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2199..2513
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2207..2225
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2243..2261
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2278..2296
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2314..2332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2350..2368
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2385..2403
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2421..2439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2486..2510
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3093..3095,3096..3809)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3135..3156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3396..3415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3743..3764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
       61 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
      121 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      181 ttttcactgc ctcgagcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      301 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      541 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      781 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      901 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      961 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1021 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1081 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1141 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1201 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1261 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1321 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1381 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1441 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1501 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1561 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1621 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1681 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1741 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1801 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1861 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1921 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1981 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2041 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2101 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     2161 aaaaataggc gtatcacgag gccctttcgt cttcactcga gtttactccc tatcagtgat
     2221 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
     2281 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     2341 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
     2401 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
     2461 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
     2521 ttcgagctcg gtacccgggg atcctctagt cagctgacgc gtgctagcgc ggccgcatcg
     2581 ataagcttac catggcaacc ctggaaaagc tgatgaaggc cttcgagtcc ctcaagtcct
     2641 tccagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     2701 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     2761 agcggcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     2821 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     2881 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcaa cagccgccac
     2941 cgccgccgcc gccgccgccg cctcctcagc ttcctcagcc gccgccgcag gcacagccgc
     3001 tgctgcctca gccgcagccg cccccgccgc cgcccccgcc gccacccggc ccggctgtgg
     3061 ctgaggagcc gctgcaccga ccgggatccc tggtgagcaa gggcgaggag ctgttcaccg
     3121 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3181 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3241 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt
     3301 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3361 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3421 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3481 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacatc agccacaacg
     3541 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     3601 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3661 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3721 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3781 ctctcggcat ggacgagctg tacaagtaat ctag
//
