LOCUS       sequence-6838-ex        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  TITLE     Unpublished, Paco
REFERENCE   2  (bases 1 to 6362)
  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"
COMMENT     Sequence Label: pRS314-URA3
FEATURES             Location/Qualifiers
     source          1..6362
                     /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        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1239..1694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1376..1397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1588..1607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1824..1846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1838..1855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1839..1855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1862..1881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1968..2771)
                     /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"
     promoter        complement(2772..2992)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     3492..3508
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3492..3508
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3542..3558)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3543..3559)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3588..3606)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3627..3643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3627..3643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3640..3662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3651..3667
                     /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(3675..3681,3682..3699,3700..3705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3858..3875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4029..4617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4788..5579,5580..5648))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5411..5430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5649..5753)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5790..6293
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6335..6353
                     /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 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg cggtgtgaaa
     1201 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt
     1261 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1321 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1381 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1441 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1501 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1561 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1621 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1681 gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1741 ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg caaggcgatt
     1801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     1861 gtaatacgac tcactatagg gcgaattggg ggtaataact gatataatta aattgaagct
     1921 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     1981 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     2041 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     2101 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     2161 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     2221 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     2281 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     2341 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     2401 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     2461 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     2521 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     2581 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     2641 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     2701 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     2761 gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag
     2821 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     2881 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     2941 agaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg aaaccccccc
     3001 cccgatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     3061 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     3121 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     3181 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     3241 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccagtgatc
     3301 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     3361 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     3421 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     3481 gcgtcggccg ctctagaact agtggatccc ccgggctgca ggaattcgat atcaagctta
     3541 tcgataccgt cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg
     3601 ttaattccga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3661 ctcacaattc cacacaacat aggagccgga agcataaagt gtaaagcctg gggtgcctaa
     3721 tgagtgaggt aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4021 ctggcgtttt tccataggct cggcccccct gacgagcatc acaaaaatcg acgctcaagt
     4081 cagaggtggc gaaacccgac aggactataa agataccagg cgttcccccc tggaagctcc
     4141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4201 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     4261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4441 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactg
     4861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5161 caacgatcaa ggcgagttac atgatccccc atgttgtgaa aaaaagcggt tagctccttc
     5221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5761 ccccgaaaag tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa
     5821 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     5881 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     5941 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     6001 gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     6061 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     6121 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     6181 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     6241 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtccta
     6301 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6361 tc
//
