LOCUS       sequence-448159-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy plasmid for inducible yeast expression of the Htt exon 1 
            construct with the last 46 amino acids of the proline-rich region at
            the N-terminus, 4 prolines left at the C-terminus using copper..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  TITLE     Herman lab plasmids 2024
REFERENCE   2  (bases 1 to 7656)
  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"
COMMENT     Sequence Label: pRS416 CUP1pro-M-PRR-Htt97QP4-YFP
FEATURES             Location/Qualifiers
     source          1..7656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..24)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8..24)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(61..81)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(61..79)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(100..116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(100..116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(113..135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    124..140
                     /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)."
     promoter        complement(join(148..154,155..172,173..178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    193..214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(331..348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(502..1090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(582..601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1261..2052,2053..2121))
                     /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     1884..1903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2122..2226)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2263..2766
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2808..2826
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2926..2945
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3439..4533)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4534..4941)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(5242..5697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5379..5400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5591..5610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5823..5845
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5837..5854
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5838..5854
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5864..5883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5864..5882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5907..5923
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5908..5924
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5959..6193
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             6689..7402
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(6728..6749)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6989..7008)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7336..7357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      7409..7656
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7425..7443)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ttttggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc
       61 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      121 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      181 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      241 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      301 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      361 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      421 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      481 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      541 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      601 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      661 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      721 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      781 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      841 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      901 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      961 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1021 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1081 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1141 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1201 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1261 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1321 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1381 cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa
     1441 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1501 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1561 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1621 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1681 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1741 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1801 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1861 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1921 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1981 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2041 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2101 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2161 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2221 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     2281 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     2341 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     2401 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     2461 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     2521 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     2581 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     2641 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     2701 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     2761 gcgatccgtc ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     2821 cgaggccctt tgtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2881 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2941 ttgtactgag agtgcaccat atcgactacg tcgttaaggc cgtttctgac agagtaaaat
     3001 tcttgaggga actttcacca ttatgggaaa tggttcaaga aggtattgac ttaaactcca
     3061 tcaaatggtc aggtcattga gtgtttttta tttgttgtat tttttttttt ttagagaaaa
     3121 tcctccaata tataaattag gaatcatagt ttcatgattt tctgttacac ctaacttttt
     3181 gtgtggtgcc ctcctccttg tcaatattaa tgttaaagtg caattctttt tccttatcac
     3241 gttgagccat tagtatcaat ttgcttacct gtattccttt acatcctcct ttttctcctt
     3301 cttgataaat gtatgtagat tgcgtatata gtttcgtcta ccctatgaac atattccatt
     3361 ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca
     3421 taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc
     3481 gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac
     3541 ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa
     3601 catcattgca gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg
     3661 agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc
     3721 caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt catcggagat
     3781 gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac
     3841 taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag ggaattcgtt
     3901 cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt
     3961 atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct
     4021 tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc
     4081 atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa
     4141 gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc
     4201 aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc
     4261 ttgttcaggt ctaacactac cggtacccca tttaggacca cccacagcac ctaacaaaac
     4321 ggcatcagcc ttcttggagg cttccagcgc ctcatctgga agtggaacac ctgtagcatc
     4381 gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc
     4441 agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc
     4501 tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata
     4561 tatatatatt gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc
     4621 acctattgga aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc
     4681 aagcatttag tcatgaacgc ttctctattc tatatgaaaa gccggttccg gcgctctcac
     4741 ctttcctttt tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa
     4801 attaacaaaa aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc
     4861 tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga
     4921 tacctgagta ttcccacagt taactgcggt caagatattt cttgaatcag gcgccttaga
     4981 ccgctcggcc aaacaaccaa ttacttgttg agaaatagag tataattatc ctataaatat
     5041 aacgtttttg aacacacatg aacaaggaag tacaggacaa ttgattttga agagaatgtg
     5101 gattttgatg taattgttgg gattccattt ttaataaggc aataatatta ggtatgtaga
     5161 tatactagaa gttctcctcg accggtcgat atgcggtgtg aaataccgca cagatgcgta
     5221 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5281 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5341 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5401 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5461 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5521 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5581 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5641 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     5701 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5761 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5821 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     5881 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     5941 attcctgcag cccccagtct agttagaaaa agacattttt gctgtcagtc actgtcaaga
     6001 gattcttttg ctggcatttc ttctagaagc aaaaagagcg atgcgtcttt tccgctgaac
     6061 cgttccagca aaaaagacta ccaacgcaat atggattgtc agaatcatat aaaagagaag
     6121 caaataactc cttgtcttgt atcaattgca ttataatatc ttcttgttag tgcaatatca
     6181 tatagaagtc atcatgcccc ctccaccccc acctcctcaa cttcctcaac ctcctccaca
     6241 ggcacagcct ctgctgcctc agccacaacc tcctccacct ccacctccac ctcctccagg
     6301 cccagctgtg gctgaggagc ctctgcaccg acctatggcg accctggaaa agctaatgaa
     6361 ggccttcgag tccctcaaaa gcttccaaca gcagcaacag caacaacagc agcaacagca
     6421 acaacagcag caacagcaac aacagcagca acagcaacaa cagcagcaac agcaacaaca
     6481 gcagcaacag caacaacagc agcaacagca acaacagcag caacagcaac aacagcagca
     6541 acagcaacaa cagcagcaac agcaacaaca gcagcaacag caacaacagc agcaacagca
     6601 acaacagcag caacagcaac aacagcagca acagcaacaa cagcagcaac agcaacaaca
     6661 gcagcaacag caacaaccgc caccacctag caagggcgag gagctgttca ccggggtggt
     6721 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     6781 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     6841 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc
     6901 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     6961 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     7021 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     7081 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     7141 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     7201 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     7261 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     7321 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     7381 catggacgag ctgtacaagt aactcgagtc atgtaattag ttatgtcacg cttacattca
     7441 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     7501 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     7561 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     7621 gagaaggttt tgggacgctc gaaggcttta atttgc
//
