LOCUS       sequence-482061-        7101 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Over-express Sc rev7 (Sc Pol zeta subunit) in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Sc rev7-pRS405/GAL
FEATURES             Location/Qualifiers
     source          1..7101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..108
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    225..246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(261..266,267..284,285..291)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    299..315
                     /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     304..326
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     323..339
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     323..339
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     358..378
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        360..378
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1378..2042
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(1391..1414)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1709..1826
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(2070..2086)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2071..2087)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2111..2130)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2112..2130)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2140..2157)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2140..2156)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2171)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2297..2752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2384..2403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2594..2615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3053..3460
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3461..4555
                     /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"
     primer_bind     complement(5049..5068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5168..5190
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5228..5246)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5314..5418
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5419..5487,5488..6279)
                     /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(5637..5656)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6450..7038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6939..6958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
       61 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      241 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      301 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      361 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctcagca
      421 gatcctgtca atgttggtca agttaaggac attcacggac gcatcccaga aatgctgtga
      481 ttatacgcaa cgatattttg cttaatttta ttttcctgtt ttatttttta ttagtggttt
      541 acagataccc tatattttat ttagttttta tacttagaga catttaattt taattccatt
      601 cttcaaattt catttttgca cttaaaacaa agatccaaaa atgctctcgc cctcttcata
      661 ttgagaatac actccattca aaattttgtc gtcaccgctg attaattttt cactaaactg
      721 atgaataatc aaaggcccca cgtcagaacc gactaaagaa gtgagtttta ttttaggagg
      781 ttgaaaacca ttattgtctg gtaaattttc atcttcttga catttaaccc agtttgaatc
      841 cctttcaatt tctgcttttt cctccaaact atcgaccctc ctgtttctgt ccaacttatg
      901 tcctagttcc aattcgatcg cattaataac tgcttcaaat gttattgtgt catcgttgac
      961 tttaggtaat ttctccaaat gcataatcaa actatttaag gaagatcgga attcgtcgaa
     1021 cacttcagtt tccgtaatga tctgatcgtc tttatccaca tgttgtaatt cactaaaatc
     1081 taaaacgtat ttttcaatgc ataaatcgtt ctttttatta ataatgcaga tggaaaatct
     1141 gtaaacgtgc gttaatttag aaagaacatc cagtataagt tcttctatat agtcaattaa
     1201 agcaggatgc ctattaatgg gaacgaactg cggcaagttg aatgactggt aagtagtgta
     1261 gtcgaatgac tgaggtgggt atacatttct ataaaataaa atcaaattaa tgtagcattt
     1321 taagtatacc ctcagccact tctctaccca tctattcata tgactagtgg atccggggtt
     1381 ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga tacttttatt
     1441 acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta aagtggttat
     1501 gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg ctgattaatt
     1561 accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt taatttgatt
     1621 cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc ataaccataa
     1681 aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca catctgcgtt
     1741 tcaggaacgc gaccggtgaa gacgaggacg cacggaggag agtcttcctt cggagggctg
     1801 tcacccgctc ggcggcttct aatccgtact tcaatatagc aatgagcagt taagcgtatt
     1861 actgaaagtt ccaaagagaa ggttttttta ggctaagata atggggctct ttacatttcc
     1921 acaacatata agtaagatta gatatggata tgtatatgga tatgtatatg gtggtaatgc
     1981 catgtaatat gattattaaa cttctttgcg tccatccaaa aaaaaagtaa gaatttttga
     2041 aaattcaagg aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     2101 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     2161 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2221 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2281 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2341 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2401 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2461 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2521 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2581 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2641 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2701 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     2761 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatcgacc ggtcgaggag
     2821 aacttctagt atatctacat acctaatatt attgccttat taaaaatgga atcccaacaa
     2881 ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt
     2941 gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag taattggttg
     3001 tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg
     3061 gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc
     3121 ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa
     3181 attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg
     3241 agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga gaagcgttca
     3301 tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt
     3361 ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc
     3421 agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat
     3481 cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct
     3541 taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg
     3601 tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa
     3661 gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt
     3721 tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag
     3781 accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc
     3841 taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag
     3901 aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt
     3961 gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat
     4021 ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga
     4081 ggaaaccatc aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc
     4141 cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat
     4201 gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc
     4261 atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg
     4321 ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc
     4381 tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga
     4441 tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag
     4501 taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga
     4561 ttctcttttt ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga
     4621 cacgaaatta caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca
     4681 tacatttatc aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata
     4741 ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag
     4801 gagggcacca cacaaaaagt taggtgtaac agaaaatcat gaaactatga ttcctaattt
     4861 atatattgga ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga
     4921 cctgaccatt tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa
     4981 agttccctca agaattttac tctgtcagaa acggccttaa cgacgtagtc gatatggtgc
     5041 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5101 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5161 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5221 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5281 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5341 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5401 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5461 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5521 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5581 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5641 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5701 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5761 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5821 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5881 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5941 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6001 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6061 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6121 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6181 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6241 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6301 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6361 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6421 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6481 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6541 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6601 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6661 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6721 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6781 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6841 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6901 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6961 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7021 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7081 tggccttttg ctcacatgtt c
//
