LOCUS       sequence-194145-        6097 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 6097)
  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 6097)
  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-Ataxin3Q22
FEATURES             Location/Qualifiers
     source          1..6097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..21
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        94..171
                     /gene="lacI"
                     /label=lacI promoter
     CDS             172..1254
                     /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(191..210)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1303..1308,1309..1326,1327..1333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1341..1357
                     /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     1365..1381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1365..1381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1377..1904
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1397..1414)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1397..1413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1406..1428)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2006..2011,2012..2027,2028..2034)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2042..2058
                     /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             2081..2734
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2692..2714
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2741..2764
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3970..3992)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4137..4155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4223..4327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4328..4396,4397..5188)
                     /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(4546..4565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5359..5947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5848..5867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
       61 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
      121 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
      181 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
      241 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
      301 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
      361 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
      421 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
      481 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
      541 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
      601 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
      661 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
      721 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
      781 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      841 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      901 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      961 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     1021 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     1081 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     1141 gtgaaaagaa aaaccacgct agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     1201 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     1261 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     1321 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     1381 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1441 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     1501 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     1561 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     1621 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     1681 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     1741 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     1801 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     1861 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1921 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1981 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     2041 attgtgagcg gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta
     2101 ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa
     2161 atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga
     2221 attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca
     2281 gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa
     2341 agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc
     2401 gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc
     2461 tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca
     2521 tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc
     2581 aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc
     2641 acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca
     2701 agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc tgttccaggg
     2761 gcccctggga tccataaaca tggagtccat cttccacgag aaacaagaag gctcactttg
     2821 tgctcaacat tgcctgaata acttattgca aggagaatat tttagccctg tggaattatc
     2881 ctcaattgca catcagctgg atgaggagga gaggatgaga atggcagaag gaggagttac
     2941 tagtgaagat tatcgcacgt ttttacagca gccttctgga aatatggatg acagtggttt
     3001 tttctctatt caggttataa gcaatgcctt gaaagtttgg ggtttagaac taatcctgtt
     3061 caacagtcca gagtatcaga ggctcaggat cgatcctata aatgaaagat catttatatg
     3121 caattataag gaacactggt ttacagttag aaaattagga aaacagtggt ttaacttgaa
     3181 ttctctcttg acgggtccag aattaatatc agatacatat cttgcacttt tcttggctca
     3241 attacaacag gaaggttatt ctatatttgt tgttaagggt gatctgccag attgcgaagc
     3301 tgaccaactc ctgcagatga ttagggtcca acagatgcat cgaccaaaac ttattggaga
     3361 agaattagca caactaaaag agcaaagagt ccataaaaca gacctggaac gagtgttaga
     3421 agcaaatgat ggctcaggaa tgttagacga agatgaggag gatttgcaga gggctctggc
     3481 actaagtcgc caagaaattg acatggaaga tgaggaagca gatctccgca gggctattca
     3541 gctaagtatg caaggtagtt ccagaaacat atctcaagat atgacacaga catcaggtac
     3601 aaatcttact tcagaagagc ttcggaagag acgagaagcc tactttgaaa aacagcagca
     3661 aaagcagcaa cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
     3721 gcagcagcag ggggacctat caggacagag ttcacatcca tgtgaaaggc cagccaccag
     3781 ttcaggagca cttgggagtg atctaggtga tgctatgagt gaagaagaca tgcttcaggc
     3841 agctgtgacc atgtctttag aaactgtcag aaatgatttg aaaacagaag gaaaaaaata
     3901 aataagaatg cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga
     3961 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4021 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4081 agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct
     4141 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4201 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4261 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4321 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4381 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4441 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4501 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4561 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4621 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4681 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4741 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4801 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4861 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4921 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4981 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5041 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5101 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5161 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5461 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6061 gtgagctgat accgctcgcc gcagccgaac gaccgag
//
