LOCUS       sequence-133467-        4879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN/ARS yeast shuttle vector with DS2 selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4879)
  AUTHORS   Rugbjerg P, Myling-Petersen N, Sommer MO
  TITLE     Flexible metabolic pathway construction using modular and divisible 
            selection gene regulators.
  JOURNAL   Metab Eng. 2015 Sep;31:189-97. doi: 10.1016/j.ymben.2015.08.004. 
            Epub 2015 Aug 21.
  PUBMED    26303342
REFERENCE   2  (bases 1 to 4879)
  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: "Metab 
            Eng."; date: "2015-09"; volume: "31:189-97. doi"; pages: " 
            10.1016/j.ymben.2015.08.004. Epub 2015 Aug 21"
COMMENT     Sequence Label: pDS2
FEATURES             Location/Qualifiers
     source          1..4879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..32)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(186..774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(266..285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(945..1736,1737..1805))
                     /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     1568..1587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1806..1910)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1947..2450
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2492..2510
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2548..2570)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2670..2689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2728..2761
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3167..3187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3194..3445,3446..3799)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAPRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     3737..3758
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     protein_bind    3812..3836
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4681..4714
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
       61 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      601 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
      661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1921 cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat aatttaaatt
     1981 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
     2041 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
     2101 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
     2161 gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
     2221 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
     2281 ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta
     2341 ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt
     2401 attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtcctaaga
     2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2701 ccataacagc tttcgacaac ccttaatata acttcgtata atgtatgcta tacgaagtta
     2761 ttaggtctag agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
     2821 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
     2881 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
     2941 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
     3001 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
     3061 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
     3121 aggatacagt tctcacatca catccgaaca taaacaagaa catatgccca agaagaagcg
     3181 gaaggtctcg agcatgaaag cgttaacggc caggcaacaa gaggtgtttg atctcatccg
     3241 tgatcacatc agccagacag gtatgccgcc gacgcgtgcg gaaatcgcgc cgcgtttggg
     3301 gttccgttcc ccaaacgcgg ctgaagaaca tctgaaggcg ctggcacgca aaggcgttat
     3361 tgaaattgtt tccggcgcat cacgcgggat tcgtctgttg caggaagagg aagaagggtt
     3421 gccgctggta ggtcgtgtgg ctgccggtga accacttctg gcgcaacagc atattgaagg
     3481 tcattatcag gtcgatcctt ccttattcaa gccgaatgct gatttcctgc tgcgcgtcag
     3541 cgggatgtcg atgaaagata tcggcattat ggatggtgac ttgctggcag tgcataaaac
     3601 tcaggatgta cgtaacggtc aggtcgttgt cgcacgtatt gatgacgaag ttaccgttaa
     3661 gcgcctgaaa aaacagggca ataaagtcga actgttgcca gaaaatagcg agtttaaacc
     3721 aattgtcgtt gaccttcgtc agcagagctt caccattgaa gggctggcgg ttggggttat
     3781 tcgcaacggc gactggctga ggtcgaatca aacaagtttg tacaaaaaag caggctccgc
     3841 ggccgccccc ttcacccgtg agtacaagct agtggtcctt ggttcaggag gcgttgggaa
     3901 gtctgctctg acagttcagt ttgttcaggg aatttttgtt gaaaaatatg acccaacgat
     3961 agaagattcc tacagaaagc aagttgaagt cgattgccaa cagtgtatgc tcgaaatcct
     4021 ggatactgca gggacagagc aatttacagc aatgagggat ttgtatatga agaacggcca
     4081 aggttttgca ctagtatatt ctattacagc tcagtccacg tttaacgact tacaggacct
     4141 gagggaacag attttacggg ttaaggacac ggaagatgtt ccaatgattt tggttggcaa
     4201 taaatgtgac ctggaagatg agcgagtagt tggcaaagag cagggccaga atttagcaag
     4261 acagtggtgt aactgtgcct ttttagaatc ttctgcaaag tcaaagatca atgttaatga
     4321 gatattttat gacctggtca gacagataaa taggaaaacg ccagtggaaa agaagaagcc
     4381 taaaaagaaa tcatgtctgc tgctctagaa gcggtaactc gaggacaata aaaagattct
     4441 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     4501 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     4561 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     4621 gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctctcgag aacccttaat
     4681 ataacttcgt ataatgtatg ctatacgaag ttattaggtg atatcagatc cactagtggc
     4741 ctatgcaccc aattgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4801 cgccagctga actgcccggg caaatcagct gcattaatga atcggccaac gcgcggggag
     4861 aggcggtttg cgtattggg
//
