LOCUS       sequence-6840-ex        7875 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 7875)
  TITLE     Unpublished, Paco
REFERENCE   2  (bases 1 to 7875)
  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-hUI
FEATURES             Location/Qualifiers
     source          1..7875
                     /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"
     primer_bind     complement(4367..4383)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4368..4384)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5101..5119)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5140..5156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5140..5156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5153..5175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5164..5180
                     /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(5188..5194,5195..5212,5213..5218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5371..5388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5542..6130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6301..7092,7093..7161))
                     /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     6924..6943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7162..7266)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7303..7806
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7848..7866
                     /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 tcgatatcaa gcttatcgat accgtcgacc
     4381 tcgagttcaa gagaaaaaaa aagaaaaagc aaaaagaaaa aaggaaagcg cgcctcgttc
     4441 agaatgacac gtatagaatg atgcattacc ttgtcatctt cagtatcata ctgttcgtat
     4501 acatacttac tgacattcat aggtatacat atatacacat gtatatatat cgtatgctgc
     4561 agctttaaat aatcggtgtc actacataag aacacctttg gtggagggaa catcgttggt
     4621 accattgggc gaggtggctt ctcttatggc aaccgcaaga gccttgaacg cactctcact
     4681 acggtgatga tcattcttgc ctcgcagaca atcaacgtgg agggtaattc tgctagcctc
     4741 tgcaaagctt tcaagaaaat gcgggatcat ctcgcaagag agatctccta ctttctccct
     4801 ttgcaaacca agttcgacaa ctgcgtacgg cctgttcgaa agatctacca ccgctctgga
     4861 aagtgcctca tccaaaggcg caaatcctga tccaaacctt tttactccac gcacggcccc
     4921 tagggcctct ttaaaagctt gaccgagagc aatcccgcag tcttcagtgg tgtgatggtc
     4981 gtctatgtgt aagtcaccaa tgcactcaac gattagcgac cagccggaat gcttggccag
     5041 agcatgtatc atatggtcca ggaattcctc gagggggggc ccggtaccca gcttttgttc
     5101 cctttagtga gggttaattc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5161 aaattgttat ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc
     5221 ctggggtgcc taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt
     5281 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5521 aaaggccgcg ttgctggcgt ttttccatag gctcggcccc cctgacgagc atcacaaaaa
     5581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgttccc
     5641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5701 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5941 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6361 agttgcctga ctgcccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gaaaaaaagc
     6721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7261 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     7321 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     7381 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     7441 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     7501 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     7561 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     7621 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     7681 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     7741 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     7801 gcgatccgtc ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7861 cgaggccctt tcgtc
//
