LOCUS       sequence-352244-        7178 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 7178)
  AUTHORS   Duennwald ML, Jagadish S, Giorgini F, Muchowski PJ, Lindquist S
  TITLE     A network of protein interactions determines polyglutamine toxicity.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Jul 18. 103(29):11051-6.
  PUBMED    16832049
REFERENCE   2  (bases 1 to 7178)
  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. 2006 Jul 18. 103(29):11051-6."
COMMENT     Sequence Label: CUP Ent2 YFP p316
FEATURES             Location/Qualifiers
     source          1..7178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..74
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(89..94,95..112,113..119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..143
                     /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     132..154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     151..167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     151..167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        188..206
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     234..250
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     235..251
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        435..669
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             1899..2612
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1926..1954)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2549..2569
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2629..2648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2630..2648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2655..2672)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2655..2671)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2664..2686)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2812..3267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2899..2918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3109..3130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3398..4201)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4202..4417)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4441..4460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4560..4582
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4620..4638)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4679..5182
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5219..5323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5324..5392,5393..6184)
                     /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(5542..5561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6355..6943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6844..6863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7097..7114
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
       61 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      121 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      181 agctcgaaat taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgaggt
      241 cgacggtatc gataagcttg atatcgaatt cccattaccg acatttgggc gctatacgtg
      301 catatgttca tgtatgtatc tgtatttaaa acacttttgt attatttttc ctcatatatg
      361 tgtataggtt tatacggatg atttaattat tacttcacca ccctttattt caggctgata
      421 tcttagcctt gttactagtt agaaaaagac atttttgctg tcagtcactg tcaagagatt
      481 cttttgctgg catttcttct agaagcaaaa agagcgatgc gtcttttccg ctgaaccgtt
      541 ccagcaaaaa agactaccaa cgcaatatgg attgtcagaa tcatataaaa gagaagcaaa
      601 taactccttg tcttgtatca attgcattat aatatcttct tgttagtgca atatcatata
      661 gaagtcatcg aaatagatat taagaaaaac aaactgtaca atcaatcaat caatcaggat
      721 ccatgaagca attgcaggaa ctacagagat tacagaagca acaacagtct ctgtctcaat
      781 ttcaagctcc tttacaacaa caacaaccac aacaacaacc agcgtactac gacattttcg
      841 gtaatccaat ctcccaagat gaatacttac agtatcagta ccaacaggac caggaacaag
      901 caatggctca gcaaagatgg ctggaccagc agcaagaaca acagcagctt gctgaacaac
      961 aatattttca gcagcaacaa caagctgcgg ccgccgcttc tgccttgcaa cagcaacaaa
     1021 cagccgctaa tatgcaacaa caacaacaac agcccgctga ttttcaacaa cctttgccta
     1081 caggttctaa taatccgttt tccatggata atcttgaaag acaaaagcag gagcaacagc
     1141 atgctcaatt gcaaagacaa caagaagaag ctagacaaca acaagaacaa ttgaagctac
     1201 aacaattgca aagacaacaa caagaggaag ctcaattaca ccagaagagg caagaagaag
     1261 cccaattaca acagcagcaa gcccaattgc tacaacagca agcccagttc cagcaacaac
     1321 aacccttgaa gcaaacaagg actgggaacc agtctatatc ggataaatac agcgacttga
     1381 ataccttgtt agcaactggt acagggatag atacttttgg taacactgga gaggcacgta
     1441 ttcctgcaca acatacaaag acaggcacat ttataaattc tcagggtaca ggctacaaac
     1501 aggttactaa tgaacccaag aacaaccctt tcttaagcaa ccaatacact ggtttaccaa
     1561 gcacaaatat cgtgcccacg caaacagggt acgggtttgg taaccaacct caaagtcctc
     1621 ctactaattc tcctcagcaa aatcctactg gtataagcta ctctcagcca caacagcaac
     1681 aacagccaca gcaacaaccg caatacatgc aaaatttcca acaacagcaa cctcaatacg
     1741 cccaaaactt ccaacaacaa ccacaataca ctcaaaatta tcaacaacaa ccacaataca
     1801 ttcaacctca tcaacaacaa cagcagcagc agcagcagca acagcagcaa cagggatata
     1861 ctcctgacca aggtgtaagc ttaattgatc ttccgcggag taaaggagaa gaacttttca
     1921 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     1981 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
     2041 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtctaa
     2101 aatgctttgc tagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc
     2161 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     2221 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     2281 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
     2341 atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac
     2401 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     2461 gcgatggccc tgtcctttta ccagacaacc attacctgtc ctatcaatct gccctttcga
     2521 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     2581 ttacacatgg catggatgaa ctatacaaat agtaagagct ccaattcgcc ctatagtgag
     2641 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2701 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2761 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     2821 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2881 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2941 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3001 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3061 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3121 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3181 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3241 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3301 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     3361 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     3421 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     3481 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     3541 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     3601 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     3661 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     3721 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     3781 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     3841 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     3901 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     3961 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     4021 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     4081 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     4141 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     4201 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     4261 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     4321 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     4381 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     4441 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4501 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4561 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4621 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgga
     4681 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     4741 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     4801 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     4861 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     4921 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     4981 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     5041 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt
     5101 aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat
     5161 aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg
     5221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5341 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5401 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5461 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5521 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5581 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5641 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5701 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5761 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5821 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5881 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5941 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6001 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6061 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6121 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6181 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6301 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6361 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6421 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6481 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6541 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6601 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6661 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6721 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6781 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6841 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6901 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6961 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7021 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7081 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7141 accgcctctc cccgcgcgtt ggccgattca ttaatgca
//
