LOCUS       sequence-83380-e        3950 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 3950)
  AUTHORS   Ito-Harashima S, McCusker JH
  TITLE     Positive and negative selection LYS5MX gene replacement cassettes 
            for use in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2004 Jan 15;21(1):53-61.
  PUBMED    14745782
REFERENCE   2  (bases 1 to 3950)
  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: "2004-01-15"; volume: "21(1)"; pages: "53-61"
COMMENT     Sequence Label: pSA39
FEATURES             Location/Qualifiers
     source          1..3950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1283..1480
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1587..1606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1588..1606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1693..1710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1864..2452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1944..1963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2623..3414,3415..3483))
                     /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     3246..3265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3484..3588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3656..3674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3712..3734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3834..3853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3934..3950,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3934..3950,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgttta gcttgccttg tccccgccgg gtcacccggc cagcgacatg
      121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      421 ggatacagtt ctcacatcac atccgaacat aaacaaccat ggttaaaacg actgaagtag
      481 taagcgaagt ttcaaaggtg gcaggtgtaa gaccatgggc aggtatattc gttgttgaaa
      541 ttcaagagga tatactcgcg gatgagttta cgttcgaggc attaatgaga actttgccat
      601 tggcgtctca agccagaatc ctcaataaaa aatcgtttca cgatagatgt tcaaatctat
      661 gcagccagct gctgcagttg tttggctgct ctatagtaac gggcttaaat tttcaagagc
      721 tgaaatttga caagggcagc ttcggtaagc cattcttaga caacaatcgt tttcttccat
      781 ttagcatgac catcggtgaa caatatgtag ctatgttcct cgtaaaatgt gtaagtacag
      841 atgaatacca ggatgtcgga attgatatcg cttctccgtg caattatggc gggagggaag
      901 agttggagct atttaaagaa gtttttagtg aaagagaatt taacggttta ctgaaagcgt
      961 ctgatccatg cacaatattt acttacttat ggtccttgaa ggagtcgtat acaaaattta
     1021 ctggaactgg ccttaacaca gacttgtcac taatagattt tggcgctatc agcttttttc
     1081 cggctgaggg agcttctatg tgcataactc tggatgaagt tccattgatt ttccattctc
     1141 aatggttcaa taacgaaatt gtcactatct gtatgccaaa gtccatcagt gataaaatca
     1201 acacgaacag accaaaatta tataatatca gcttatctac gttgattgat tatttcatcg
     1261 aaaatgatgg tttataatca gtactgacaa taaaaagatt cttgttttca agaacttgtc
     1321 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     1381 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     1441 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     1501 ccgccatcca gtgtcgaaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     1561 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     1621 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3661 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3721 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3781 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3841 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
     3901 ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
