LOCUS       sequence-122355-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  URA+ yeast shuttle vector containing yZ3EV TF cassette and a Z3pr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  TITLE     pRS416-yZ3EV-Z3pr
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7793)
  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            "Unpublished"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: pRS416-yZ3EV-Z3pr (RB3578)
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2003..2250)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2216..2234
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2277..2300)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(5004..5024)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5004..5022)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5043..5059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5043..5059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5056..5078)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5067..5083
                     /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(5091..5097,5098..5115,5116..5121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5136..5157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5274..5291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5445..6033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5525..5544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6204..6995,6996..7064))
                     /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     6827..6846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7065..7169)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7206..7709
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7750..7768
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg ccgcaaatta aagccttcga gcgtcccaaa accttctcaa
     2041 gcaaggtttt cagtataatg ttacatgcgt acacgcgtct gtacagaaaa aaaagaaaaa
     2101 tttgaaatat aaataacgtt cttaatacta acataactat aaaaaaataa atagggacct
     2161 agacttcagg ttgtctaact ccttcctttt cggttagagc ggatgtgggg ggagggcgtg
     2221 aatgtaagcg tgacataact aattacatga aaactcgagt atagtttttt ctccttgacg
     2281 ttaaagtata gaggtatatt aacaattttt tgttgatact tttatgacat ttgaataaga
     2341 agtaatacaa actgaaaatg ttgaaagtat tagttaaagt ggttatgcag cttttccatt
     2401 tatatatctg ttaatagatc aaaaatcatc gcttcgctga ttaattaccc cagaaataag
     2461 gctaaaaaac taatcgcatt atcatcctat ggttgttaat ttgattcgtt aatttgaagg
     2521 tttgtggggc caggttactg ccaatttttc ctcttcataa ccataaaagc tagtattgta
     2581 gaatctttat tgttcggagc agtgcggcgc gaggcacatc tgcgtttcag gaacgcgacc
     2641 ggtgaagacg aggacgcacg gtctagacgc ccacgcacgc ccacgcccgc ccacgcacgc
     2701 ccacgcccgc ccacgcacgc ccacgcggcc gcacttcaat atagcaatga gcagttaagc
     2761 gtattactga aagttccaaa gagaaggttt ttttaggcta agataatggg gctctttaca
     2821 tttccacaac atataagtaa gattagatat ggatatgtat atggatatgt atatggtggt
     2881 aatgccatgt aatatgatta ttaaacttct ttgcgtccat ccaaaaaaaa agtaagaatt
     2941 tttgaaaatt caatataaac tagtcccttt aaaaacatat gcctcaccct aacatatttt
     3001 ccaattaacc ctcaatattt ctctgtcacc cggcctctat tttccatttt cttctttacc
     3061 cgccacgcgt ttttttcttt caaatttttt tcttccttct tctttttctt ccacgtcctc
     3121 ttgcataaat aaataaaccg ttttgaaacc aaactcgcct ctctctctcc tttttgaaat
     3181 atttttgggt ttgtttgatc ctttccttcc caatctctct tgtttaatat atattcattt
     3241 atatcacgct ctctttttat cttccttttt ttcctctctc ttgtattctt ccttcccctt
     3301 tctactcaaa ccaagaagaa aaagaaaagg tcaatctttg ttaaagaata ggatcttcta
     3361 ctacatcagc ttttagattt ttcacgctta ctgctttttt cttcccaaga tcgaaaattt
     3421 actgaattaa caatgggaac gagaccctac gcttgccctg tagaaagttg tgataggaga
     3481 ttcagcagat ctgacgagct gactaggcat attcgtattc acaccggcca aaagcctttc
     3541 caatgtagaa tctgtatgag aaacttcagc agaagtgacc atttgactac gcacattaga
     3601 acccacaccg gcgagaagcc gttcgcgtgt gacatttgtg gaagaaagtt cgccagatct
     3661 gatgaaagaa agagacatac aaagatccat acaggaggtg ggggcactcc ggcagcagcc
     3721 tcaaccctag aagatccaag cgctggagac atgagggcag ccaacttgtg gccaagccca
     3781 ttaatgatca aaagatctaa gaagaattct cttgcactgt ctttgaccgc tgatcaaatg
     3841 gtttccgcac tacttgatgc tgaaccacct atattgtaca gtgaatacga cccaactcgt
     3901 cctttttccg aggcttccat gatgggtctt ttgacaaatc tagcagacag ggaattagtt
     3961 catatgatta attgggccaa aagagttccc ggctttgttg acttgacatt acatgaccag
     4021 gtacatcttc ttgaatgcgc gtggttagaa atattgatga tcggattagt gtggagaagt
     4081 atggagcacc ctgtaaaatt attatttgca cctaatttat tattggacag gaaccaaggt
     4141 aaatgcgtag aaggtatggt tgagattttt gatatgcttc ttgccactag ctccaggttc
     4201 agaatgatga atttgcaagg agaagaattc gtttgcctga aatcaattat tttgttaaat
     4261 agcggtgtct atacattttt aagctcaact ctgaaaagcc tggaagagaa agaccatata
     4321 catagagtgt tggacaagat cactgataca ttgattcacc ttatggcgaa agcaggattg
     4381 actctgcagc aacagcatca gaggctagct cagttactac taattttgtc acacattaga
     4441 catatgtcta acaagggtat ggagcacctg tattctatga agtgtaaaaa cgtcgttcct
     4501 ctgtatgatt tattactaga aatgctggac gctcacaggt tgcatgcacc aacttctaga
     4561 ggcggagcct ccgttgaaga aaccgaccaa tcccacctag cgacagcagg gtctacatct
     4621 agcgaactgc atttggacgg tgaagacgtt gccatggccc acgctgacgc tttagacgat
     4681 tttgatttag atatgttggg tgacggggac tctccagggc ctggcttcac tccacatgat
     4741 tcagcacctt acggtgcgct ggatatggct gacttcgaat ttgagcagat gttcaccgat
     4801 gctctgggta ttgatgagta tggtggttag tagtaagctg gttcttcttt ctgccattta
     4861 attgtagact tgttttctat ttaatatcac tgtatcctgt agcgtttgta tacattccgt
     4921 aatagaaggt ccgatacttt actttcatat cagtcgcagt ttgccacttg aggggagatc
     4981 gacgcgagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     5041 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5221 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5641 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5881 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7081 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7141 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc
     7201 ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa atatataaat
     7261 taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt
     7321 aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca
     7381 ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt
     7441 gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc
     7501 taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt
     7561 tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa
     7621 aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg
     7681 aggcgcgtgt aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta
     7741 acctataaaa ataggcgtat cacgaggccc tttcgtcacg aggccctttc gtc
//
