LOCUS       sequence-82749-e        6934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration cassette to create p1-Short-UL*(TGA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6934)
  AUTHORS   Ravikumar A, Arrieta A, Liu CC
  TITLE     An orthogonal DNA replication system in yeast.
  JOURNAL   Nat Chem Biol. 2014 Mar;10(3):175-7. doi: 10.1038/nchembio.1439. 
            Epub 2014 Feb 2.
  PUBMED    24487693
REFERENCE   2  (bases 1 to 6934)
  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"; doi: 
            "10.1038/nchembio.1439"; journalName: "Nat Chem Biol"; date: 
            "2014-03"; volume: "10"; issue: "3"; pages: "175-7"
COMMENT     Sequence Label: pCCL-GKL1ShortMutPRC-Ug1Lg2starTGA
FEATURES             Location/Qualifiers
     source          1..6934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1995..2798
                     /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="MHKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(2947..4041)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MFVPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMAL*HEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4358..4376)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4372..4419
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4486..4577
                     /gene="bla"
                     /label=AmpR promoter
     terminator      5442..5481
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      5512..6100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6001..6020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6254..6271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6286..6425
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6351..6370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6511..6533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6527..6718)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctcgagagta ctaaatttag tcctttgtca aaattttctg atgttaaaaa tataacagta
       61 tgttttgata tagaatctta ttttgatcca gaaaaagaat ctaatcaagt taatataccc
      121 tttatatgtt gtgcatctat aatatataat aaagtcatag gaaatattgt agattttgaa
      181 ggaagagatt gtgtagctca aatgatagaa tatgttgtag atatatgtgg agagcttaat
      241 atatcttcag tggaactaat tgcacataat ggtggaggtt atgattttca ttatatttta
      301 agtagtatgt ataatcctgc agctattaaa aatatattaa ttagaaataa ctcatttata
      361 agttttaatt ttgctcacga tggagtcaaa ttttctgtaa aagattccta tagtttcttg
      421 ttatgtagtt tagcaaatgc ttcaaaagca tttttaaacg aagaaacctt taagaaaaca
      481 gattttcccc atcatgattt aaaaacagca gatgatttat ataaagtata taaagaatgg
      541 tcatctgtaa acactgaaat aaatcatgta gtggaaaaag aaaaacttct tataacatca
      601 gaacatatag ttaatttcac taaaaatgat aaatctaaaa ctctaataga atggtctaaa
      661 gattattgta gaaatgatgt tttggtttta tctaaggtat ggttagaatt taaaaatgct
      721 gtagaagata tttttaattg tgaattagta gatcaaacta tgacattagc aggactaagt
      781 tataaattat ttcaagcaaa tatgcctttt gatgttgaat taagacatcc aaataaagaa
      841 gattatttta acatgagaga ggctttaata ggagggagat gtattagtgt caatggaata
      901 tataaagatg ttttatgttt agatgtaaaa tcattatatc cagcatctat ggcattttat
      961 gaccagccat atggatcttt caaaagagta tctagtagac ctaaagatga attaggtatt
     1021 tattatgtca gagtaactcc taatagaaat aataaatcca acttttttcc tataagaagt
     1081 cacaataaaa ttacttataa taattttgaa gaaagtacat atatagcatg gtatacaaat
     1141 gtagatatag atataggttt gtctgaaggt cataatatag aatatatccc ctttgattct
     1201 tatggaaata taggttattc ttggtctaaa aaaggtaaaa tattcgaaaa atatataaaa
     1261 gacgtgctgt acaaattaaa aataaagtat gaaaaacaaa acaataaagt taaaagaaat
     1321 gttatcaaaa ttattatgaa cagtttatgg ggcaaattcg cacaaaaatg ggtaaatttt
     1381 gagtatttta taaaatcaga agatgatata gattttgagt cagaagaggc atataagata
     1441 tgggacactg attttatgct gataaagaaa attaaagaat ctacttattc atctaaacct
     1501 atacaaaatg gagtatttac attaagttgg gcaagatacc acatgaaaag tatatgggat
     1561 gcaggggcta aagaaggagc agaatgtatc tattcggaca cagatagtat ttttgtacat
     1621 aaagaacatt ttaaaaagaa tgctaaattt atgttaaatg gtttaaaagt tcctattata
     1681 ggatcagaag taggacaatt agaattagaa tgtgagtttg ataaattgtt atgtgcaggt
     1741 aaaaagcaat acatgggatt ttatacttat tttcaagatg gaaaaccatg tataaaagaa
     1801 aagaaaagat ttaagggtat tcctagtaat tatataatac ctgaattata tgctcattta
     1861 ctttcaggtg cagacaaaga agctaaaata caatttttga aatttagaag agaatgggga
     1921 tcagttaaag gatatataga aaataagacc gtgaaagcta cttaatatat gaaagttttt
     1981 ataataatta taaaatgcat aaagctacat ataaggaacg tgctgctact catcctagtc
     2041 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     2101 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     2161 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     2221 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
     2281 acattggtaa tacagtcaaa ttgcagtatt ctgcgggtgt atacagaata gcagaatggg
     2341 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     2401 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     2461 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     2521 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     2581 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     2641 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     2701 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     2761 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     2821 caggtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagc
     2881 tccgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga
     2941 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttcggtggt
     3001 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
     3061 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
     3121 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg
     3181 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
     3241 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
     3301 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
     3361 ggcagaatca atcaattgat gttgaacctt caatgtaggg aattcgttct tgatggtttc
     3421 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
     3481 aataggcaat ggtggctcat gtcatagggc catgaaagcg gccattcttg tgattctttg
     3541 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
     3601 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
     3661 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
     3721 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
     3781 aacactaccg gtaccccatt taggaccacc cacagcacct aacaaaacgg catcagcctt
     3841 cttggaggct tccagcgcct catctggaag tggaacacct gtagcatcga tagcagcacc
     3901 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
     3961 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
     4021 gatcttctta ggaacaaaca tatgcaattt gtttgtcaaa atatctattt tgttctttta
     4081 atgtatttat tatactttca atattgtttt cacaattaag atctttagct atctcactag
     4141 tgttcacatt agaaattaat tttcctcttt tagaaaattt attaacttct ccatctataa
     4201 atgatatacg taatttttta tctatacaag atcggatccc ctcagactca gattggtgta
     4261 aagatcctca aggcaataga ataacagatt ttgatagtac ttctagaaga tccggctgct
     4321 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     4381 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     4441 gactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
     4501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4561 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4621 cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
     4681 aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
     4741 ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
     4801 gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
     4861 attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
     4921 tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
     4981 tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
     5041 accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
     5101 agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
     5161 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
     5221 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
     5281 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
     5341 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
     5401 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
     5461 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
     5521 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5581 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5641 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5701 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5761 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5821 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5881 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5941 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6001 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6061 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6121 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6181 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6241 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     6301 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     6361 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     6421 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6481 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6541 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     6601 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     6661 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     6721 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
     6781 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     6841 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     6901 gcgcttcgtt aatacagatg taggtgttgg cacc
//
