LOCUS       sequence-448235-        6441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy plasmid for inducible yeast expression of the Htt exon 1 
            construct without the last 36 amino acids of the proline-rich region
            (135-170) tagged with YFP using copper..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  TITLE     Herman lab plasmids 2024
REFERENCE   2  (bases 1 to 6441)
  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-Htt103QdeltaP135-170-YFP
FEATURES             Location/Qualifiers
     source          1..6441
                     /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(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     2807..2825
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2925..2944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2968..3183
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3184..3987
                     /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"
     rep_origin      complement(4118..4573)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4255..4276)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4467..4486
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4699..4721
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4713..4730
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4714..4730
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4740..4759
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4740..4758
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4783..4799
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4784..4800
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4835..5069
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             5454..6167
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(5493..5514)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5754..5773)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6101..6122
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      6174..6421
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6190..6208)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6422..6438)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     2821 gaggcccttt gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2881 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     2941 tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta ttcttttttt
     3001 tgatttcggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     3061 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     3121 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     3181 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     3241 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     3301 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     3361 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     3421 ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat
     3481 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     3541 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     3601 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     3661 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     3721 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     3781 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     3841 gtggatgatg tggtgtctac aggatctgac attattattg ttggaagagg actatttgca
     3901 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     3961 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     4021 aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc ggtgtgaaat
     4081 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     4141 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4201 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4261 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4321 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4381 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4441 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4501 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4561 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4621 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4681 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     4741 aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg
     4801 ataagcttga tatcgaattc ctgcagcccc cagtctagtt agaaaaagac atttttgctg
     4861 tcagtcactg tcaagagatt cttttgctgg catttcttct agaagcaaaa agagcgatgc
     4921 gtcttttccg ctgaaccgtt ccagcaaaaa agactaccaa cgcaatatgg attgtcagaa
     4981 tcatataaaa gagaagcaaa taactccttg tcttgtatca attgcattat aatatcttct
     5041 tgttagtgca atatcatata gaagtcatca tggcgaccct ggaaaagcta atgaaggcct
     5101 tcgagtccct caaaagcttc caacagcagc aacagcaaca acagcagcaa cagcaacaac
     5161 agcagcaaca gcaacaacag cagcaacagc aacaacagca gcaacagcaa caacagcagc
     5221 aacagcaaca acagcagcaa cagcaacaac agcagcaaca gcaacaacag cagcaacagc
     5281 aacaacagca gcaacagcaa caacagcagc aacagcaaca acagcagcaa cagcaacaac
     5341 agcagcaaca gcaacaacag cagcaacagc aacaacagca gcaacagcaa caacagcagc
     5401 aacagcaaca accgccacca cctccccctc cacccccacc tcctcaactt cctagcaagg
     5461 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     5521 gccacaggtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     5581 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     5641 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     5701 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     5761 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     5821 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     5881 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     5941 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     6001 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     6061 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     6121 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaactc gagtcatgta
     6181 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
     6241 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     6301 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     6361 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     6421 cgatccacta gttctagagc g
//
