LOCUS       sequence-83375-e        4727 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 4727)
  AUTHORS   Goldstein AL, McCusker JH
  TITLE     Three new dominant drug resistance cassettes for gene disruption in 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1999 Oct;15(14):1541-53.
  PUBMED    10514571
REFERENCE   2  (bases 1 to 4727)
  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: "1999-10"; volume: "15(14)"; pages: "1541-53"
COMMENT     Sequence Label: pAG35
FEATURES             Location/Qualifiers
     source          1..4727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            572..1690
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        572..915
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             916..1488
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDTEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHACRRMGFT
                     LCGLDTALYDGTASDGEQALYMGMPCP"
     terminator      1493..1690
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2364..2383)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2365..2383)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2470..2487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2641..3229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2721..2740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3400..4191,4192..4260))
                     /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     4023..4042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4261..4365)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4433..4451
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4489..4511)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4611..4630
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4711..4727,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4711..4727,1)
                     /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 caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt
      961 gtcccggggg acaccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtc
     1021 ttccgcgtca ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc
     1081 ctgaccaagg tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc
     1141 gacccggact cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg
     1201 gtcgtctcgt actccggctg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg
     1261 gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgcacgc
     1321 gagcggggcg ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac
     1381 gcgtgccggc ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc
     1441 gcctcggacg gcgagcaggc gctctacatg ggcatgccct gcccctaaca gtactgacaa
     1501 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     1561 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     1621 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc attcgtatgt gaatgctggt
     1681 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgacta gggttgctgc
     1741 catcggcctc gctcgcgtct ttgccggata gcaagagcgc ctttggcctc tacgagccct
     1801 gccacggctc tgcgcccgat ctgcccgccg ggaaggcgaa cccgatcgga tgcatcctct
     1861 ctgctgccat gatgctgaag ttgtcgttga acatggttgc tgccggcgag gcggtcgagc
     1921 aggcagtgca ggaggtgttg gactcgggag tcagaacggg cgacctgctc ggctcgagct
     1981 ccacttcgga ggttggcgac gccattgcgc ttgcagttaa ggaagccttg cgcgggcaat
     2041 ccgcagctgg tctgagctag cctcgaggac tcttctcttt agactattct actcttatgc
     2101 acgtaaaaaa ttctaggaaa tatgtattaa ctagaagtaa aataaccggc tagtggcatt
     2161 catatagccg tctgtttaca tctacatcac acatttcgag tgtatatctc gcaacgttgg
     2221 cgttaaatag gcagtcaatg gcccgaccat tctatggtgt ttaggtcgat gccatctttg
     2281 tacgtttaaa cggtaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     2341 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     2401 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2461 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2521 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2581 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2641 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2701 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2761 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2821 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2881 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2941 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3001 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3061 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3121 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3181 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3241 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3301 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3361 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3421 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3481 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3541 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3601 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3661 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3721 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3781 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3841 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3901 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3961 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4021 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4081 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4141 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4201 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4261 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4381 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4441 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4501 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4561 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4621 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     4681 atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
