LOCUS       sequence-83383-e        5138 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 5138)
  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 5138)
  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: pSA54
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        572..915
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1911..2108
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2775..2794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2776..2794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2881..2898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3052..3640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3132..3151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3811..4602,4603..4671))
                     /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     4434..4453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4672..4776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4844..4862
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4900..4922)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5022..5041
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5122..5138,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5122..5138,1)
                     /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 caaccatgac attacaagat catcttaatg ataaatcaaa tatcatccta
      961 cttttcacca ctagattaac taatgaatta attgaatttt ggaatgatga ttataatttt
     1021 gaatgttctt taagattatt aaatgattta tcattacaac aaaaaatatc ttcaattaca
     1081 aaaaacccca accatcaaca acaaagattt aaatataaac aattattatc aaatttattt
     1141 attagatatg caatcaattc aatcattcaa tcaaatgatc tatttaaaaa aattgaattt
     1201 gattataatg aatttgggaa acccatatta ccatataatt tccaattcaa tattagtagt
     1261 tctaatgata taattgcctt aattgtggaa tttccatctt catcttcatt atcatcatca
     1321 tcattgtcat catcattatc tggtgatcga gcttatgacc caattggtat tgatttatca
     1381 cattctattc aaaatagtat atcaaataca gaatatcttg aacaatttaa accgattttt
     1441 gatgatgttt atgaattacc acaaattgat aattatttta aatttaatca tttttggaca
     1501 ttaaaagaat catttacaaa attgattggt agtggattaa atattgaatt atcagatttt
     1561 tatttcatag ttgattataa tgataatgag ttttctagtg ataatgaaaa attccaaaca
     1621 agtatagttg gaaataatat tggagatagt tggatgtatt attcattgaa ttggtttgat
     1681 aaaatcttga taaatataga taaacttgaa ttagccaaga atcaatttat aacggcatta
     1741 aataaaaaag aattttattg tcgtagttct attcttgata atggtaatgt cgatgaatct
     1801 aaattgccag taattataac gataattaat caaaacaaga ataaatcaat tcaaccaata
     1861 gagttgagat ttgaaaaatt attatctgaa catttaaacc agtgatcagt actgacaata
     1921 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     1981 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     2041 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     2101 ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgactagg gttgctgcca
     2161 tcggcctcgc tcgcgtcttt gccggatagc aagagcgcct ttggcctcta cgagccctgc
     2221 cacggctctg cgcccgatct gcccgccggg aaggcgaacc cgatcggatg catcctctct
     2281 gctgccatga tgctgaagtt gtcgttgaac atggttgctg ccggcgaggc ggtcgagcag
     2341 gcagtgcagg aggtgttgga ctcgggagtc agaacgggcg acctgctcgg ctcgagctcc
     2401 acttcggagg ttggcgacgc cattgcgctt gcagttaagg aagccttgcg caggcaatcc
     2461 gcagctggtc tgagctagcc tcgaggaccc ttctctttag actattctac tcttatgcac
     2521 gtaaaaaatt ctaggaaata tgtattaact aggagtaaaa taaccggcta gtggcattca
     2581 tatagccgtc tgtttacatc tacatcacac atttcgagtg tatatctcgc aacgttggcg
     2641 ttaaataggc agtcaatggc ccgaccattc tatggtgttt aggtcgatgc catctttgta
     2701 cgtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     2761 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     2821 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2881 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2941 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3241 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3481 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3601 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3661 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3721 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3781 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3841 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3901 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3961 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4021 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4081 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4141 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4201 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4261 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4321 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4381 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4441 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4501 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4561 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4621 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4681 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4741 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4801 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4861 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4921 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4981 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5041 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     5101 cttatgtatc atacacatac gatttaggtg acactata
//
