LOCUS       sequence-83384-e        4018 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 4018)
  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 4018)
  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: pSA40
FEATURES             Location/Qualifiers
     source          1..4018
                     /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      1309..1506
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1569..1602
                     /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(1655..1674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1656..1674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1761..1778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1932..2520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2012..2031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2691..3482,3483..3551))
                     /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     3314..3333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3552..3656)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3724..3742
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3780..3802)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3902..3921
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4002..4018,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4002..4018,1)
                     /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 aaccatggtt aaaacgactg aagtagtaag cgaagtttca aaggtggcag gtgtaagacc
      541 atgggcaggt atattcgttg ttgaaattca agaggatata ctcgcggatg agtttacgtt
      601 cgaggcatta atgagaactt tgccattggc gtctcaagcc agaatcctca ataaaaaatc
      661 gtttcacgat agatgttcaa atctatgcag ccagctgctg cagttgtttg gctgctctat
      721 agtaacgggc ttaaattttc aagagctgaa atttgacaag ggcagcttcg gtaagccatt
      781 cttagacaac aatcgttttc ttccatttag catgaccatc ggtgaacaat atgtagctat
      841 gttcctcgta aaatgtgtaa gtacagatga ataccaggat gtcggaattg atatcgcttc
      901 tccgtgcaat tatggcggga gggaagagtt ggagctattt aaagaagttt ttagtgaaag
      961 agaatttaac ggtttactga aagcgtctga tccatgcaca atatttactt acttatggtc
     1021 cttgaaggag tcgtatacaa aatttactgg aactggcctt aacacagact tgtcactaat
     1081 agattttggc gctatcagct tttttccggc tgagggagct tctatgtgca taactctgga
     1141 tgaagttcca ttgattttcc attctcaatg gttcaataac gaaattgtca ctatctgtat
     1201 gccaaagtcc atcagtgata aaatcaacac gaacagacca aaattatata atatcagctt
     1261 atctacgttg attgattatt tcatcgaaaa tgatggttta taatcagtac tgacaataaa
     1321 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     1381 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     1441 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     1501 atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctctcgagaa
     1561 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtgat atcagatcca
     1621 ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg
     1681 ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1741 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2101 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     2161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2341 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     2401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3541 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3661 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3721 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3781 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3841 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     3901 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc
     3961 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactata
//
