LOCUS       sequence-475694-        7297 bp    DNA     circular SYN 11-JUN-2025
DEFINITION  Overexpress Sc RFC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7297)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Sc RFC[2+5]-pRS403/GAL
FEATURES             Location/Qualifiers
     source          1..7297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     309..326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    443..464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(479..484,485..502,503..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    517..533
                     /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     522..544
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     541..557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     541..557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     576..596
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        578..596
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1731..2395
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(1744..1767)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    2062..2179
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(3565..3584)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3566..3584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3594..3611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3594..3610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3603..3625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3751..4206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3838..3857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4048..4069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4466..5128)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5129..5316)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5463..5482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5582..5604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5642..5660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5728..5832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5833..5901,5902..6693)
                     /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(6051..6070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6864..7297,1..155)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
       61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      181 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      241 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      301 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      541 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      601 aaaagctgga gcttacttta tggatcctct atattcaatt atataaagat gcctgtatat
      661 tagagttggg atattttaac caataagttt aataattgga tgtgctcgtt cgttccgttg
      721 tttaatctgg aatccgtagt aaacagcaac caagatattt gatttttgaa gttcgtatca
      781 aagttatcgt tggtgatgta atattcatgt agttggttaa ctactgacgc agcagaccaa
      841 ccgcttttca taaacgtatt gacgtatttc ttaatctcat caaaatcacc acttttgact
      901 ttttcaacaa tttctattag aatgtcatga ggtaccacac cagctaattc ttccacttgg
      961 gtagaagtta tgtttttacc atcacccaag tattgcgccc ccttggaagc ggattgtaat
     1021 aatgtaatgc ctcttctcaa gtcacctgcc gaaatgtcca agatcctttc caaaactcca
     1081 tcatcacatt taacattttc ttgttcgctg ataaacctca aacggtcaat ggcattactt
     1141 gcgtctaggg ctttgaatct aaatttggaa caacgggacg ctagcgggtc aataattctt
     1201 gtcacataat tacaaatcaa acagaatctt gttactcctg aataggtttc cattgttcta
     1261 cgtaatgcac tctgtgcatc ggcagtcatt gaatcggcct catcgagaat aatgatctta
     1321 tacggtgggc acggatagtt ctccaagtca tgtttgctag gcttggatac agtcaatctt
     1381 gcaaaatttt ttaccttttc tcttacaata gagataccac gttcgtcaga agcgttcaac
     1441 tccaatattc ttgatttcat caagtcgggt ccatataatt cttttgttag cgctaaaatg
     1501 gtagacgttt taccagttcc tgggggacca taaaacaaca tatgtggtag attagctgac
     1561 tttaaggttt tcttcaaaac agtaacggca tgatcttgag ctgtcacttc atctaggttt
     1621 ttgggcctgt atttctcaac ccagggttgt tgtgccaatg attgctctgc ggctaacttt
     1681 gatatcttac gctttttatt tggaccaaac ccttcaaaca tggatccggg gttttttctc
     1741 cttgacgtta aagtatagag gtatattaac aattttttgt tgatactttt attacatttg
     1801 aataagaagt aatacaaacc gaaaatgttg aaagtattag ttaaagtggt tatgcagttt
     1861 ttgcatttat atatctgtta atagatcaaa aatcatcgct tcgctgatta attaccccag
     1921 aaataaggct aaaaaactaa tcgcattatc atcctatggt tgttaatttg attcgttcat
     1981 ttgaaggttt gtggggccag gttactgcca atttttcctc ttcataacca taaaagctag
     2041 tattgtagaa tctttattgt tcggagcagt gcggcgcgag gcacatctgc gtttcaggaa
     2101 cgcgaccggt gaagacgagg acgcacggag gagagtcttc cttcggaggg ctgtcacccg
     2161 ctcggcggct tctaatccgt acttcaatat agcaatgagc agttaagcgt attactgaaa
     2221 gttccaaaga gaaggttttt ttaggctaag ataatggggc tctttacatt tccacaacat
     2281 ataagtaaga ttagatatgg atatgtatat ggatatgtat atggtggtaa tgccatgtaa
     2341 tatgattatt aaacttcttt gcgtccatcc aaaaaaaaag taagaatttt tgaaaattca
     2401 aggaattcat atgtcattgt gggtagataa atacagacct aagtccttga atgctctttc
     2461 acataatgaa gagttgacaa attttctaaa atcgttatct gatcagcctc gtgatttacc
     2521 tcatctttta ctgtatggac caaatggtac aggtaagaaa acgcgttgta tggcattatt
     2581 ggagtccata tttggacctg gagtctatag attgaaaatt gatgtcagac aatttgtcac
     2641 tgcttcgaac agaaaactag aactgaatgt ggtcagctcg ccataccatt tagagatcac
     2701 gccaagtgat atgggtaaca atgatagaat tgtcatccaa gaactattga aagaagtggc
     2761 tcaaatggaa caagtggact ttcaagattc taaggatgga cttgcccata gatataagtg
     2821 tgttattatc aacgaggcga actcgttaac aaaagatgct caagctgctt taagacgtac
     2881 catggaaaaa tactccaaaa acattaggtt gataatggtc tgcgattcga tgtcgcctat
     2941 aattgctcct atcaaatccc gttgtctgtt gattcgttgt cctgcaccaa gcgatagcga
     3001 aatttcaact atcttgtctg atgtggtgac aaatgaaaga atacaactag aaacaaagga
     3061 tattttaaaa agaattgctc aggcatcgaa tggaaacttg cgagtctccc tattaatgct
     3121 tgaatctatg gcactgaaca acgaattagc attgaaaagc agtagcccta taataaaacc
     3181 agattggatt atagtgatcc ataaattaac gaggaaaatc gttaaagaga gatctgtcaa
     3241 ttctttaatc gaatgcagag ctgtcctata cgatttacta gctcattgta tacctgccaa
     3301 tatcatctta aaggaactaa cgttttcttt gttggatgtg gaaaccctga ataccacgaa
     3361 taaatcgtcc ataattgaat actcaagtgt ttttgacgaa agattatcac ttggaaacaa
     3421 agcaatattc catttggaag ggttcatagc aaaagttatg tgctgtctag attaatgtaa
     3481 gatatgtcat aaatactgta taagtcacac aaaaagcgga tctgctgagc atcgatctcg
     3541 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     3601 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3661 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3721 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     3781 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3841 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3901 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3961 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4021 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4081 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4141 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4201 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga
     4261 tccgtcgagt tcaagagaaa aaaaaagaaa aagcaaaaag aaaaaaggaa agcgcgcctc
     4321 gttcagaatg acacgtatag aatgatgcat taccttgtca tcttcagtat catactgttc
     4381 gtatacatac ttactgacat tcataggtat acatatatac acatgtatat atatcgtatg
     4441 ctgcagcttt aaataatcgg tgtcactaca taagaacacc tttggtggag ggaacatcgt
     4501 tggtaccatt gggcgaggtg gcttctctta tggcaaccgc aagagccttg aacgcactct
     4561 cactacggtg atgatcattc ttgcctcgca gacaatcaac gtggagggta attctgctag
     4621 cctctgcaaa gctttcaaga aaatgcggga tcatctcgca agagagatct cctactttct
     4681 ccctttgcaa accaagttcg acaactgcgt acggcctgtt cgaaagatct accaccgctc
     4741 tggaaagtgc ctcatccaaa ggcgcaaatc ctgatccaaa cctttttact ccacgcacgg
     4801 cccctagggc ctctttaaaa gcttgaccga gagcaatccc gcagtcttca gtggtgtgat
     4861 ggtcgtctat gtgtaagtca ccaatgcact caacgattag cgaccagccg gaatgcttgg
     4921 ccagagcatg tatcatatgg tccagaaacc ctatacctgt gtggacgtta atcacttgcg
     4981 attgtgtggc ctgttctgct actgcttctg cctctttttc tgggaagatc gagtgctcta
     5041 tcgctagggg accacccttt aaagagatcg caatctgaat cttggtttca tttgtaatac
     5101 gctttactag ggctttctgc tctgtcatct ttgccttcgt ttatcttgcc tgctcatttt
     5161 ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca ttatgtgata
     5221 atgccaatcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa aaatgaaaat
     5281 cattaccgag gcataaaaaa atatagagtg tactagagga ggccaagagt aatagaaaaa
     5341 gaaaattgcg ggaaaggact gtgttatgac ttccctgact aatgccgtgt tcaaacgata
     5401 cctggcagtg actcctagcg ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc
     5461 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5521 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5581 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5641 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5701 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6061 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6841 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6901 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6961 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7021 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7081 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7141 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7201 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7261 gaaagcgcca cgcttcccga agggagaaag gcggaca
//
