LOCUS       sequence-217945-        5753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cwc25 in pRS316(Ura) vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5753)
  TITLE     Cwc25 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5753)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pRS316-Cwc25
FEATURES             Location/Qualifiers
     source          1..5753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..80)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     180..199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        223..439
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             440..1243
                     /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(1374..1829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1511..1532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1723..1742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1955..1977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1969..1986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1970..1986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1993..2012
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1993..2011
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2020..2127
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2044..2060
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2044..2060
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2094..2110)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2095..2111)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2139..2157)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2178..2194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2178..2194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2191..2213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2202..2218
                     /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(2226..2232,2233..2250,2251..2256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2271..2292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2409..2426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2660..2679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3339..4130,4131..4199))
                     /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     3962..3981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4341..4844
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4885..4903
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcttggttcc tcgtcggata gtggcaagtt cgcgcgtttc ggtgatgacg gtgaaaacct
       61 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      181 ggcatcagag cagattgtac tgagagtgca ccataccacc ttttcaattc atcatttttt
      241 tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga
      301 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
      361 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
      421 caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc
      481 tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc
      541 ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa
      601 aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt
      661 taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt
      721 tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga
      781 atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa
      841 gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc
      901 atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag
      961 cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg
     1021 ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg
     1081 tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg
     1141 aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc
     1201 aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt
     1261 aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     1321 ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     1381 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1441 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1501 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1561 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1621 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1681 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1741 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1801 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     1861 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1921 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1981 gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg cggtggcggc
     2041 cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc
     2101 gtcgacctcg agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg
     2161 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2221 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2281 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2341 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4321 gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa
     4381 tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt
     4441 gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg
     4501 ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac
     4561 acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa
     4621 tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa
     4681 acctttgttt attttttttt cttcattccg taactcttct accttcttta tttactttct
     4741 aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca
     4801 tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcta agaaaccatt
     4861 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcgttcaagg
     4921 cttacgccga gtcgcttact cgctgctcgc tttctttctt agtgttgtta tcctatataa
     4981 aacgtaactg tttagttatt taagtccaag tgcatattta cactataata tataatataa
     5041 catctcatct ttttttattt ctttttcttc ttctcccgcc gcgggcttac atacagagat
     5101 ataagaacaa gtggtacgta gcatgcagct tctcaaaaga gatgagcgtt cagttttgag
     5161 tacagggaag aaaggccaaa tcgagagtac attatcatgg ggtcgggcga tttgaatctg
     5221 ttgaaatcat ggaatcccaa gcttatgaaa aatagaaaaa aagtatggga gaccgaacag
     5281 gatctaatta ccgaacagca gaagcttaac acgcgattga aagaaattga aaaagaacga
     5341 gaactaaatg aactgctgaa cgaatcgagc aaggataagc cggagactct caagaatgac
     5401 ttggctttga agaagtcggg cttggaatgg atgtatcagg atgcaaaact gagtgacgaa
     5461 aaggaagact atttattagg taagaaaaaa ctagattcat ctatactaaa ccaacctgca
     5521 acaccgcctg taagggcggc cactactatt tccgcttctg gtgccgcaac atccataagt
     5581 tcgcagaaga agaagtcaaa attgcttaaa gatgatccaa tgagtaagtt taaggtaact
     5641 aaacaacaaa gaagaactcc agactcaaca aaaaaaagag caatgtcgca acgagggaaa
     5701 ccgttgtcca agcccgctcc ggacctagac tactaaactt tccttttctt aga
//
