LOCUS       sequence-83391-e        6951 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 6951)
  AUTHORS   Vorachek-Warren MK, McCusker JH
  TITLE     DsdA (D-serine deaminase): a new heterologous MX cassette for gene 
            disruption and selection in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2004 Jan 30;21(2):163-71.
  PUBMED    14755641
REFERENCE   2  (bases 1 to 6951)
  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: "Yeast."; 
            date: "2004-01-30"; volume: "21(2)"; pages: "163-71"
COMMENT     Sequence Label: pMKV006
FEATURES             Location/Qualifiers
     source          1..6951
                     /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        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2057..2090
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2145..2488
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3823..4020
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4083..4116
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4177..4195)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4216..4232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4216..4232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4229..4251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4240..4256
                     /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(4264..4270,4271..4288,4289..4294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4447..4464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4618..5206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5377..6168,6169..6237))
                     /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     6000..6019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6238..6342)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6379..6882
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6924..6942
                     /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 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcca ccgcggtggc ggccgccagc tgaagcttcg tacgctgcag
     2041 gtcgacaacc cttaatataa cttcgtataa tgtatgctat acgaagttat taggtctaga
     2101 gatctgttta gcttgccttg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     2161 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     2221 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     2281 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     2341 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     2401 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     2461 ctcacatcac atccgaacat aaacaaccat ggaaaacgct aaaatgaact cgctcatcgc
     2521 ccagtatccg ttggtaaagg atctggttgc tcttaaagaa accacctggt ttaatcctgg
     2581 cacgacctca ttggctgaag gtttacctta tgttggcctg accgaacagg atgttcagga
     2641 cgcccatgcg cgcttatccc gttttgcacc ctatctggca aaagcatttc ctgaaactgc
     2701 tgccactggg gggattattg aatcagaact ggttgccatt ccagctatgc aaaaacggct
     2761 ggaaaaagaa tatcagcaac cgatcagcgg gcaactgtta ctgaaaaaag atagccattt
     2821 gcccatttcc ggctccataa aagcacgcgg cgggatttat gaagtcctgg cacacgcaga
     2881 aaaactggct ctggaagcgg ggttgctgac gcttgatgat gactacagca aactgctttc
     2941 tccggagttt aaacagttct ttagccaata cagcattgct gtgggctcaa ccggaaatct
     3001 ggggttatca atcggcatta tgagcgcccg cattggcttt aaggtgacag ttcatatgtc
     3061 tgctgatgcc cgggcatgga aaaaagcgaa actgcgcagc catggcgtta cggtcgtgga
     3121 atatgagcaa gattatggtg ttgccgtcga ggaaggacgt aaagcagcgc agtctgaccc
     3181 gaactgtttc tttattgatg acgaaaattc ccgcacgttg ttccttgggt attccgtcgc
     3241 tggccagcgt cttaaagcgc aatttgccca gcaaggccgt atcgtcgatg ctgataaccc
     3301 tctgtttgtc tatctgccgt gtggtgttgg cggtggtcct ggtggcgtcg cattcgggct
     3361 taaactggcg tttggcgatc atgttcactg cttttttgcc gaaccaacgc actccccttg
     3421 tatgttgtta ggcgtccata caggattaca cgatcagatt tctgttcagg atattggtat
     3481 cgacaacctt accgcagcgg atggccttgc agttggtcgc gcatcaggct ttgtcgggcg
     3541 ggcaatggag cgtctgctgg atggcttcta tacccttagc gatcaaacca tgtatgacat
     3601 gcttggctgg ctggcgcagg aagaaggtat tcgtcttgaa ccttcggcac tggcgggtat
     3661 ggccggacct cagcgcgtgt gtgcatcagt aagttaccaa cagatgcacg gtttcagcgc
     3721 agaacaactg cgtaatacca ctcatctggt gtgggcgacg ggaggtggaa tggtgccgga
     3781 agaagagatg aatcaatatc tggcaaaagg ccgttaatca gtactgacaa taaaaagatt
     3841 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     3901 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     3961 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     4021 ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctctcg agaaccctta
     4081 atataacttc gtataatgta tgctatacga agttattagg tgatatcaga tccactagtg
     4141 gcctatgcgg ccgcccgctc tagccagctt ttgttccctt tagtgagggt taattccgag
     4201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4261 acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta
     4321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4621 ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc tcgtgcgctc
     4741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4801 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5041 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc ccgtcgtgta
     5461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5761 gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat
     5821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6361 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     6421 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     6481 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     6541 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     6601 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     6661 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     6721 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     6781 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     6841 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa gaaaccatta
     6901 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
