LOCUS       sequence-83381-e        4121 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 4121)
  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 4121)
  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: pSA50
FEATURES             Location/Qualifiers
     source          1..4121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1454..1651
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1758..1777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1759..1777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1864..1881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2035..2623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2115..2134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2794..3585,3586..3654))
                     /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     3417..3436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3655..3759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3827..3845
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3883..3905)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4005..4024
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4105..4121,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4105..4121,1)
                     /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 gacattacaa gatcatctta
      481 atgataaatc aaatatcatc ctacttttca ccactagatt aactaatgaa ttaattgaat
      541 tttggaatga tgattataat tttgaatgtt ctttaagatt attaaatgat ttatcattac
      601 aacaaaaaat atcttcaatt acaaaaaacc ccaaccatca acaacaaaga tttaaatata
      661 aacaattatt atcaaattta tttattagat atgcaatcaa ttcaatcatt caatcaaatg
      721 atctatttaa aaaaattgaa tttgattata atgaatttgg gaaacccata ttaccatata
      781 atttccaatt caatattagt agttctaatg atataattgc cttaattgtg gaatttccat
      841 cttcatcttc attatcatca tcatcattgt catcatcatt atctggtgat cgagcttatg
      901 acccaattgg tattgattta tcacattcta ttcaaaatag tatatcaaat acagaatatc
      961 ttgaacaatt taaaccgatt tttgatgatg tttatgaatt accacaaatt gataattatt
     1021 ttaaatttaa tcatttttgg acattaaaag aatcatttac aaaattgatt ggtagtggat
     1081 taaatattga attatcagat ttttatttca tagttgatta taatgataat gagttttcta
     1141 gtgataatga aaaattccaa acaagtatag ttggaaataa tattggagat agttggatgt
     1201 attattcatt gaattggttt gataaaatct tgataaatat agataaactt gaattagcca
     1261 agaatcaatt tataacggca ttaaataaaa aagaatttta ttgtcgtagt tctattcttg
     1321 ataatggtaa tgtcgatgaa tctaaattgc cagtaattat aacgataatt aatcaaaaca
     1381 agaataaatc aattcaacca atagagttga gatttgaaaa attattatct gaacatttaa
     1441 accagtgatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1501 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1561 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1621 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     1681 agtgtcgaaa acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     1741 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     1801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2221 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2461 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3781 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3841 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3901 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3961 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4021 tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat
     4081 aaccttatgt atcatacaca tacgatttag gtgacactat a
//
