LOCUS       sequence-448243-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy plasmid for inducible yeast expression of the Htt exon 1 
            construct without the first 10 amino acids of the proline-rich 
            region (125-134) tagged with YFP using copper. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  TITLE     Herman lab plasmids 2024
REFERENCE   2  (bases 1 to 6572)
  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-Htt103QdeltaP125-134-YFP
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..21)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5..21)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(58..78)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(58..76)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(97..113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(97..113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(145..151,152..169,170..175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    190..211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(328..345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(499..1087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(579..598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1258..2049,2050..2118))
                     /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     1881..1900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2119..2223)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2260..2763
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2804..2822
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2860..2882)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2982..3001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3025..3240
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3241..4044
                     /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(4175..4630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4524..4543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4756..4778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4770..4787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4771..4787
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4797..4816
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4797..4815
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4840..4856
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4841..4857
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4887..5121
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             5605..6318
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(5644..5665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5905..5924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6252..6273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      6325..6572
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6341..6359)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 gggggatcca ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct
       61 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      121 ttgttattcg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      181 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      241 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      301 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      361 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      421 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      901 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1141 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1201 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1561 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1861 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2221 gcgcacattt ccccgaaaag tgccacctgg gtccttttca tcacgtgcta taaaaataat
     2281 tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat
     2341 taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca
     2401 aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct
     2461 tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat
     2521 cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct
     2581 ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta
     2641 ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt
     2701 aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg
     2761 atccgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2821 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2881 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2941 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     3001 actgagagtg caccatacca ccttttcaat tcatcatttt ttttttattc ttttttttga
     3061 tttcggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     3121 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     3181 tgcccagtat tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc
     3241 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     3301 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     3361 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     3421 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     3481 tccgccaagt acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca
     3541 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     3601 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     3661 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     3721 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     3781 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     3841 acacccggtg tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg
     3901 gatgatgtgg tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     3961 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     4021 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     4081 tcacaaatta gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc
     4141 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     4201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4621 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4681 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4741 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     4801 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     4861 agcttgatat cgaattcctg cagcccctag ttagaaaaag acatttttgc tgtcagtcac
     4921 tgtcaagaga ttcttttgct ggcatttctt ctagaagcaa aaagagcgat gcgtcttttc
     4981 cgctgaaccg ttccagcaaa aaagactacc aacgcaatat ggattgtcag aatcatataa
     5041 aagagaagca aataactcct tgtcttgtat caattgcatt ataatatctt cttgttagtg
     5101 caatatcata tagaagtcat catggcgacc ctggaaaagc taatgaaggc cttcgagtcc
     5161 ctcaaaagct tccaacagca gcaacagcaa caacagcagc aacagcaaca acagcagcaa
     5221 cagcaacaac agcagcaaca gcaacaacag cagcaacagc aacaacagca gcaacagcaa
     5281 caacagcagc aacagcaaca acagcagcaa cagcaacaac agcagcaaca gcaacaacag
     5341 cagcaacagc aacaacagca gcaacagcaa caacagcagc aacagcaaca acagcagcaa
     5401 cagcaacaac agcagcaaca gcaacaacag cagcaacagc aacaacagca gcaacagcaa
     5461 caaccgccac cacctcaacc tcctccacag gcacagcctc tgctgcctca gccacaacct
     5521 cctccacctc cacctccacc tcctccaggc ccagctgtgg ctgaggagcc tctgcaccga
     5581 cctctagaac tagagcttat cgatagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     5641 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgaggga
     5701 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     5761 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     5821 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     5881 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5941 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     6001 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     6061 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     6121 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     6181 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     6241 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     6301 gacgagctgt acaagtaact cgagtcatgt aattagttat gtcacgctta cattcacgcc
     6361 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     6421 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     6481 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     6541 aggttttggg acgctcgaag gctttaattt gc
//
