LOCUS       sequence-83385-e        4189 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 4189)
  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 4189)
  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: pSA53
FEATURES             Location/Qualifiers
     source          1..4189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1480..1677
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1740..1773
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1826..1845)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1827..1845)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1932..1949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2183..2202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2862..3653,3654..3722))
                     /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     3485..3504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3723..3827)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3895..3913
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3951..3973)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4073..4092
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4173..4189,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4173..4189,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc tgtttagctt gccttgtccc
      121 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
      181 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      241 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      301 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      361 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      421 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      481 aaccatgaca ttacaagatc atcttaatga taaatcaaat atcatcctac ttttcaccac
      541 tagattaact aatgaattaa ttgaattttg gaatgatgat tataattttg aatgttcttt
      601 aagattatta aatgatttat cattacaaca aaaaatatct tcaattacaa aaaaccccaa
      661 ccatcaacaa caaagattta aatataaaca attattatca aatttattta ttagatatgc
      721 aatcaattca atcattcaat caaatgatct atttaaaaaa attgaatttg attataatga
      781 atttgggaaa cccatattac catataattt ccaattcaat attagtagtt ctaatgatat
      841 aattgcctta attgtggaat ttccatcttc atcttcatta tcatcatcat cattgtcatc
      901 atcattatct ggtgatcgag cttatgaccc aattggtatt gatttatcac attctattca
      961 aaatagtata tcaaatacag aatatcttga acaatttaaa ccgatttttg atgatgttta
     1021 tgaattacca caaattgata attattttaa atttaatcat ttttggacat taaaagaatc
     1081 atttacaaaa ttgattggta gtggattaaa tattgaatta tcagattttt atttcatagt
     1141 tgattataat gataatgagt tttctagtga taatgaaaaa ttccaaacaa gtatagttgg
     1201 aaataatatt ggagatagtt ggatgtatta ttcattgaat tggtttgata aaatcttgat
     1261 aaatatagat aaacttgaat tagccaagaa tcaatttata acggcattaa ataaaaaaga
     1321 attttattgt cgtagttcta ttcttgataa tggtaatgtc gatgaatcta aattgccagt
     1381 aattataacg ataattaatc aaaacaagaa taaatcaatt caaccaatag agttgagatt
     1441 tgaaaaatta ttatctgaac atttaaacca gtgatcagta ctgacaataa aaagattctt
     1501 gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca
     1561 aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta
     1621 agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg
     1681 tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctctcgaga acccttaata
     1741 taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc actagtggcc
     1801 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     1861 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2281 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2521 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3841 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3901 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3961 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4021 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4081 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     4141 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
