LOCUS       sequence-6839-ex        7836 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 7836)
  TITLE     Unpublished, Paco
REFERENCE   2  (bases 1 to 7836)
  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-hUD
FEATURES             Location/Qualifiers
     source          1..7836
                     /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"
     promoter        1908..2095
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        3221..3441
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3442..4245
                     /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(5062..5080)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5101..5117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5101..5117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5114..5136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5125..5141
                     /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(5149..5155,5156..5173,5174..5179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5332..5349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5503..6091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6262..7053,7054..7122))
                     /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     6885..6904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7123..7227)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7264..7767
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7809..7827
                     /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 gcgaattgga gctcgcccgg ggagcctcta gtacactcta
     1921 tattttttta tgcctcggta atgattttca tttttttttt tccacctagc ggatgactct
     1981 ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat aaagtaatgt
     2041 gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg caaagatgac
     2101 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
     2161 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
     2221 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
     2281 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
     2341 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
     2401 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
     2461 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
     2521 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
     2581 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
     2641 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagaga
     2701 agccacctcg cccaatgggt ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg
     2761 acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg
     2821 cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa
     2881 ggatcgctcg cggctcttac cagcctaact tcgatcactg gaccgctgat cgtcacggcg
     2941 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta
     3001 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga
     3061 atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat
     3121 tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg
     3181 ccatctccag cagccgcacg cggcgcatcg gggggggggt ttcaattcaa ttcatcattt
     3241 tttttttatt cttttttttg atttcggttt ctttgaaatt tttttgattc ggtaatctcc
     3301 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     3361 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc
     3421 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     3481 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     3541 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     3601 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     3661 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa
     3721 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     3781 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     3841 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     3901 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     3961 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     4021 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg
     4081 ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt
     4141 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     4201 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     4261 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     4321 cccctagtgg atcccccggg ctgcaggaat tcctggacca tatgatacat gctctggcca
     4381 agcattccgg ctggtcgcta atcgttgagt gcattggtga cttacacata gacgaccatc
     4441 acaccactga agactgcggg attgctctcg gtcaagcttt taaagaggcc ctaggggccg
     4501 tgcgtggagt aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag
     4561 cggtggtaga tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga
     4621 aagtaggaga tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta
     4681 gcagaattac cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga
     4741 gtgcgttcaa ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg
     4801 atgttccctc caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat
     4861 acgatatata tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa
     4921 cagtatgata ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag
     4981 gcgcgctttc cttttttctt tttgcttttt cttttttttt ctcttgaact cgaggggggg
     5041 cccggtaccc agcttttgtt ccctttagtg agggttaatt ccgagcttgg cgtaatcatg
     5101 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc
     5161 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     5221 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5281 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5341 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5401 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctcggccc
     5521 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5581 ataaagatac caggcgttcc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5641 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5701 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5761 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5821 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5881 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5941 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6001 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6061 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6121 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6181 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6241 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6301 ctgtctattt cgttcatcca tagttgcctg actgcccgtc gtgtagataa ctacgatacg
     6361 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6421 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6481 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6541 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6601 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6661 ccccatgttg tgaaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6721 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6781 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6841 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6901 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6961 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7021 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7081 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7141 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7201 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt
     7261 ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat atataaatta
     7321 aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa
     7381 aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg
     7441 tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga
     7501 ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta
     7561 ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt
     7621 tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa
     7681 cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag
     7741 gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac cattattatc atgacattaa
     7801 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
