LOCUS       sequence-83396-e        4845 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 4845)
  AUTHORS   Hartzog PE, Nicholson BP, McCusker JH
  TITLE     Cytosine deaminase MX cassettes as positive/negative selectable 
            markers in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2005 Jul 30;22(10):789-98.
  PUBMED    16088873
REFERENCE   2  (bases 1 to 4845)
  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: "2005-07-30"; volume: "22(10)"; pages: "789-98"
COMMENT     Sequence Label: pBN42
FEATURES             Location/Qualifiers
     source          1..4845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            565..1921
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        565..908
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             909..1718
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(976..995)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1724..1921
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2482..2501)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2483..2501)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2588..2605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2759..3347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2839..2858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3518..4309,4310..4378))
                     /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     4141..4160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4379..4483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4551..4569
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4607..4629)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4729..4748
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4829..4845,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4829..4845,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctcatta ttgcactggg tgttctgggc gggactgcac acgctgccaa
      121 tgaagtaact ttcctgggtt cagtaagcgc aacaacctgt gacctgacca cttctgtcaa
      181 cggcgccgct cagccgaacc aggttgttca gctgggtacc gttcaggcta accagccggg
      241 taattttgtt gacttcgcca tgaaaccggt tgacccgaat gcacagggtt gtgcaaatct
      301 ggctcagaaa acggccactg tttcctgggc ttctgctgcc ctggatggcg aaggctttgg
      361 ggcaaccagc ggtacagcta ctgatgccaa agtactggtt gaaagcgtga actccaaaaa
      421 cccgggggct gtaaacgcta acgccagcac ggttgacttt gaaggtgcca agctgaccac
      481 cgacggcctg cagttcaaag ccaagctgaa aggtggtgcg accgagttta gcttgcctcg
      541 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
      601 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
      661 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
      721 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
      781 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
      841 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
      901 aaacaaccat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat tccaacatgg
      961 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1021 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1081 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1141 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1201 cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1261 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1321 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     1381 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1441 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     1501 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     1561 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     1621 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     1681 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agtactgaca ataaaaagat
     1741 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     1801 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     1861 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     1921 gctgtcgatt cgatactaac gccgccatcc agtgtcgaca ttattgcact gggtgttctg
     1981 ggcgggactg cacacgctgc caatgaagta actttcctgg gttcagtaag cgcaacaacc
     2041 tgtgacctga ccacttctgt caacggcgcc gctcagccga accaggttgt tcagctgggt
     2101 accgttcagg ctaaccagcc gggtaatttt gttgacttcg ccatgaaacc ggttgacccg
     2161 aatgcacagg gttgtgcaaa tctggctcag aaaacggcca ctgtttcctg ggcttctgct
     2221 gccctggatg gcgaaggctt tggggcaacc agcggtacag ctactgatgc caaagtactg
     2281 gttgaaagcg tgaactccaa aaacccgggg gctgtaaacg ctaacgccag cacggttgac
     2341 tttgaaggtg ccaagctgac caccgacggc ctgcagttca aagccaagct gaaaggtggt
     2401 gcgaccgagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2461 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2521 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2941 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3181 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4381 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4441 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4501 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4561 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4621 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4681 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4741 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4801 acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
