LOCUS       sequence-483376-        7152 bp    DNA     circular SYN 14-AUG-2025
DEFINITION  Over-express Sc Pol12 (Sc pol alpha subunit) in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Sc Pol12-pRS406/GAL
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     608..627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(846..950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1018..1036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1074..1096)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1196..1215
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1239..1454
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1455..2258
                     /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(2389..2844)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2526..2547)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2738..2757
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2970..2992
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2984..3001
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2985..3001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3011..3030
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3011..3029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3054..3070
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3055..3071
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3099..3763
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3315..3432
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3727..3750
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3768..3784)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3784)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5937..5957)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5937..5955)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5976..5992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5976..5992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5989..6011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6000..6016
                     /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(6024..6030,6031..6048,6049..6054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6069..6090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6207..6224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7137..7152,1..776,777..845))
                     /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"
ORIGIN
        1 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
       61 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      121 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      181 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
      241 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
      301 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      361 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      421 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      481 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
      541 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
      601 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      661 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      721 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      781 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
      841 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      961 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     1021 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     1081 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1141 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     1201 tcagagcaga ttgtactgag agtgcaccat accacctttt caattcatca tttttttttt
     1261 attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     1321 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     1381 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acctgcagga
     1441 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     1501 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     1561 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     1621 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     1681 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
     1741 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     1801 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     1861 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     1921 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     1981 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     2041 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     2101 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     2161 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     2221 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     2281 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttaccctatg
     2341 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     2401 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2461 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2521 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2581 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2641 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2701 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2761 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2821 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     2881 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     2941 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     3001 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
     3061 cgacggtatc gataagcttg atatcgaatt ccttgaattt tcaaaaattc ttactttttt
     3121 tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac catatacata
     3181 tccatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
     3241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
     3301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
     3361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
     3421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
     3481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
     3541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
     3601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
     3661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
     3721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat ccactagttc
     3781 tagagcggcc gcatgagcgg atctatcgat gtcatcactc attttgggcc tgatgcagac
     3841 aagccggaaa tcatcactgc attagagaat ttaactaaac tgcatgcatt gtccgtcgag
     3901 gatctataca ttaaatggga gcagttctcc aatcaaaggc gtcaaactca cacagatctg
     3961 acttccaaga acatagatga gtttaaacaa tttttacaat tacaaatgga aaagcgtgct
     4021 aaccaaattt catcgtcctc taaggtaaac acatccacaa aaaaaccagt gattaaaaaa
     4081 agtttgaatt caagtccctt gtttggcctt agtattccaa agactccgac tttaaaaaaa
     4141 agaaagttac atggtccctt ttcactcagt gattccaagc aaacatataa tgtaggctct
     4201 gaggcagaaa caaatgaaaa aggaaatagt tctttaaaat tggagtttac tccaggcatg
     4261 gcagaagatg ctgtgggtga cagtgctcca ttgtcccacg ctaaaagctc ggatgccaag
     4321 actcccggct cgtcgacatt ccaaactcca acaacaaaca ctcctaccac atcgaggcaa
     4381 aatgttcctg ctggtgaaat tttggattct ttgaaccctg aaaatattga aatatcatct
     4441 ggtaatccaa atgtaggcct tctttcgact gaagaaccat catataacca ggtcaaagta
     4501 gaaccatttt atgatgctaa aaagtataaa ttcagaacaa tgaggcaaaa cttacaggaa
     4561 gcgtctgatg tcctcgatga tcaaattgaa tcgtttacta agattattca aaatcactat
     4621 aagttatctc caaacgactt cgctgatcca actattcaat ctcagtccga aatttatgct
     4681 gttggaagga tagttccaga ttcgccgacc tacgacaagt ttttgaatcc agaatcgctt
     4741 tccctagaga cctcaagaat gggcggcgtt ggaagaaggg taaggttaga tctatcgcag
     4801 gtcaacgagc tttcgttttt cttaggccaa attgtggcat tcaaaggtaa aaatgctaat
     4861 ggggattact tcaccgttaa ctctattttg ccgctacctt atccaaactc cccagtgtcc
     4921 acatcgcagg aactgcagga gtttcaagca aatttggaag gttcttcttt gaaggttatc
     4981 gtgacctgtg gcccatactt tgcaaatgat aacttctccc tcgaattgtt gcaagaattt
     5041 atcgatagta taaataatga ggtaaaacct cacgtcttga taatgtttgg cccatttata
     5101 gatatcaccc atcctctgat agcgagtggt aaattgccaa attttcccca gttcaagaca
     5161 cagcccaaaa cattggacga gctttttttg aagttattta ctcctattct aaaaaccata
     5221 tcaccacata tccaaaccgt attgataccc tccacaaaag atgcaatttc taatcacgcc
     5281 gcatatccac aagcttcttt aattagaaag gctttacagc tgcctaaaag gaacttcaaa
     5341 tgcatggcta atccttcatc ttttcaaata aatgagatat acttcggttg ctcaaacgtt
     5401 gatacattca aagatctaaa ggaagtcata aaaggcggta ctacctcatc aagatataga
     5461 ttagatcgtg tttctgagca tattttacaa cagcgcaggt attaccccat atttccaggt
     5521 agtattcgta caagaataaa accaaaagac gtatctacga agaaggaaac caatgatatg
     5581 gaaagcaaag aagaaaaagt ttatgaacat atatctggtg cagatttaga cgtgagttat
     5641 ctgggactaa cagagtttgt tggtgggttc tcgcctgaca taatgattat acccagtgaa
     5701 ttacaacact ttgcaagagt tgtccaaaac gtagttgtta taaatcccgg aagatttatt
     5761 agagcaacag gtaacagggg atcctatgca caaattactg tccaatgccc tgatcttgaa
     5821 gacgggaaat tgacgcttgt tgaaggtgaa gagccagttt atctacacaa cgtgtggaag
     5881 cgcgctagag ttgacttgat tgctagttga ccgcggtgga gctccagctt ttgttccctt
     5941 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6001 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6061 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6121 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7081 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7141 caatgcttaa tc
//
