LOCUS       sequence-194140-        6184 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 6184)
  AUTHORS   Winborn BJ, Travis SM, Todi SV, Scaglione KM, Xu P, Williams AJ, 
            Cohen RE, Peng J, Paulson HL
  TITLE     The deubiquitinating enzyme ataxin-3, a polyglutamine disease 
            protein, edits Lys63 linkages in mixed linkage ubiquitin chains.
  JOURNAL   J Biol Chem. 2008 Sep 26. 283(39):26436-43.
  PUBMED    18599482
REFERENCE   2  (bases 1 to 6184)
  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: "J Biol 
            Chem. 2008 Sep 26. 283(39):26436-43."
COMMENT     Sequence Label: pGEX-6P1-Ataxin3Q80
FEATURES             Location/Qualifiers
     source          1..6184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..128
                     /gene="lacI"
                     /label=lacI promoter
     CDS             129..1211
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(148..167)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1260..1265,1266..1283,1284..1290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1298..1314
                     /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     1322..1338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1322..1338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1334..1861
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1354..1371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1354..1370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1363..1385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1963..1968,1969..1984,1985..1991)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1999..2015
                     /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)."
     CDS             2038..2691
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2649..2671
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2698..2721
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4014..4036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4181..4199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4267..4371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4372..4440,4441..5232)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4590..4609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5403..5991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5892..5911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6145..6162
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
       61 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      121 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      181 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      241 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      301 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
      361 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
      421 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
      481 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
      541 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
      601 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
      661 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
      721 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
      781 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      841 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
      901 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
      961 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1021 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     1081 tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct
     1141 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1201 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
     1261 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1321 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
     1381 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1441 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1501 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
     1561 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
     1621 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
     1681 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     1741 ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg
     1801 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     1861 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     1921 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg
     1981 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg
     2041 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     2101 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     2161 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     2221 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     2281 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     2341 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     2401 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     2461 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     2521 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     2581 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     2641 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg
     2701 gaagttctgt tccaggggcc cctgggatcc ataaacatgg agtccatctt ccacgagaaa
     2761 caagaaggct cactttgtgc tcaacattgc ctgaataact tattgcaagg agaatatttt
     2821 agccctgtgg aattatcctc aattgcacat cagctggatg aggaggagag gatgagaatg
     2881 gcagaaggag gagttactag tgaagattat cgcacgtttt tacagcagcc ttctggaaat
     2941 atggatgaca gtggtttttt ctctattcag gttataagca atgccttgaa agtttggggt
     3001 ttagaactaa tcctgttcaa cagtccagag tatcagaggc tcaggatcga tcctataaat
     3061 gaaagatcat ttatatgcaa ttataaggaa cactggttta cagttagaaa attaggaaaa
     3121 cagtggttta acttgaattc tctcttgacg ggtccagaat taatatcaga tacatatctt
     3181 gcacttttct tggctcaatt acaacaggaa ggttattcta tatttgttgt taagggtgat
     3241 ctgccagatt gcgaagctga ccaactcctg cagatgatta gggtccaaca gatgcatcga
     3301 ccaaaactta ttggagaaga attagcacaa ctaaaagagc aaagagtcca taaaacagac
     3361 ctggaacgaa tgttagaagc aaatgatggc tcaggaatgt tagacgaaga tgaggaggat
     3421 ttgcagaggg ctctggcact aagtcgccaa gaaattgaca tggaagatga ggaagcagat
     3481 ctccgcaggg ctattcagct aagtatgcaa ggtagttcca gaaacatatc tcaagatatg
     3541 acacagacat caggtacaaa tcttacttca gaagagcttc ggaagagacg agaagcctac
     3601 tttgaaaaac agcagcaaaa gcagcaacag cagcagcagc agcagcagca gcagcagcag
     3661 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     3721 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcgggac
     3781 ctatcaggac agagttcaca tccatgtgaa aggccagcca ccagttcagg agcacttggg
     3841 agtgatctag gtgatgctat gagtgaagaa gacatgcttc aggcagctgt gaccatgtct
     3901 ttagaaactg tcagaaatga tttgaaaaca gaaggaaaaa aataaataag aatgcggccg
     3961 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     4021 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4081 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     4141 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     4201 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4261 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4321 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4381 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4441 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4501 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4561 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4621 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4681 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4741 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4801 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4861 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4921 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4981 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5041 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5101 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5161 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5221 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5281 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5341 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5401 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5461 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5521 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5581 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5641 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5701 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5761 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5821 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5881 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5941 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6001 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6061 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6121 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     6181 atgc
//
