LOCUS       sequence-83382-e        4967 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 4967)
  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 4967)
  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: pSA48
FEATURES             Location/Qualifiers
     source          1..4967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        572..915
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1740..1937
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2604..2623)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2605..2623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2710..2727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2881..3469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2961..2980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3640..4431,4432..4500))
                     /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     4263..4282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4501..4605)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4673..4691
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4729..4751)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4851..4870
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4951..4967,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4951..4967,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgcccg ccgggaaggc gaacccgatc ggatgcatcc tctctgctgc
      121 catgatgctg aagttgtcgt tgaacatggt tgctgccggc gaggcggtcg agcaggcagt
      181 gcaggaggtg ttggactcgg gagtcagaac gggcgacctg ctcggctcga gctccacttc
      241 ggaggttggc gacgccattg cgcttgcagt taaggaagcc ttgcgcaggc aatccgcagc
      301 tggtctgagc tagcctcgag gacccttctc tttagactat tctactctta tgcacgtaaa
      361 aaattctagg aaatatgtat taactaggag taaaataacc ggctagtggc attcatatag
      421 ccgtctgttt acatctacat cacacatttc gagtgtatat ctcgcaacgt tggcgttaaa
      481 taggcagtca atggcccgac cattctatgg tgtttaggtc gatgccatct ttgtacagct
      541 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
      601 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
      661 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
      721 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
      781 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
      841 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
      901 cgaacataaa caaccatggt taaaacgact gaagtagtaa gcgaagtttc aaaggtggca
      961 ggtgtaagac catgggcagg tatattcgtt gttgaaattc aagaggatat actcgcggat
     1021 gagtttacgt tcgaggcatt aatgagaact ttgccattgg cgtctcaagc cagaatcctc
     1081 aataaaaaat cgtttcacga tagatgttca aatctatgca gccagctgct gcagttgttt
     1141 ggctgctcta tagtaacggg cttaaatttt caagagctga aatttgacaa gggcagcttc
     1201 ggtaagccat tcttagacaa caatcgtttt cttccattta gcatgaccat cggtgaacaa
     1261 tatgtagcta tgttcctcgt aaaatgtgta agtacagatg aataccagga tgtcggaatt
     1321 gatatcgctt ctccgtgcaa ttatggcggg agggaagagt tggagctatt taaagaagtt
     1381 tttagtgaaa gagaatttaa cggtttactg aaagcgtctg atccatgcac aatatttact
     1441 tacttatggt ccttgaagga gtcgtataca aaatttactg gaactggcct taacacagac
     1501 ttgtcactaa tagattttgg cgctatcagc ttttttccgg ctgagggagc ttctatgtgc
     1561 ataactctgg atgaagttcc attgattttc cattctcaat ggttcaataa cgaaattgtc
     1621 actatctgta tgccaaagtc catcagtgat aaaatcaaca cgaacagacc aaaattatat
     1681 aatatcagct tatctacgtt gattgattat ttcatcgaaa atgatggttt ataatcagta
     1741 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     1801 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     1861 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     1921 tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg tcgactaggg
     1981 ttgctgccat cggcctcgct cgcgtctttg ccggatagca agagcgcctt tggcctctac
     2041 gagccctgcc acggctctgc gcccgatctg cccgccggga aggcgaaccc gatcggatgc
     2101 atcctctctg ctgccatgat gctgaagttg tcgttgaaca tggttgctgc cggcgaggcg
     2161 gtcgagcagg cagtgcagga ggtgttggac tcgggagtca gaacgggcga cctgctcggc
     2221 tcgagctcca cttcggaggt tggcgacgcc attgcgcttg cagttaagga agccttgcgc
     2281 aggcaatccg cagctggtct gagctagcct cgaggaccct tctctttaga ctattctact
     2341 cttatgcacg taaaaaattc taggaaatat gtattaacta ggagtaaaat aaccggctag
     2401 tggcattcat atagccgtct gtttacatct acatcacaca tttcgagtgt atatctcgca
     2461 acgttggcgt taaataggca gtcaatggcc cgaccattct atggtgttta ggtcgatgcc
     2521 atctttgtac gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     2581 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     2641 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3061 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3301 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3481 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3541 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4621 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4681 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4741 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4801 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4861 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     4921 atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
