LOCUS       sequence-83370-e        4880 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 4880)
  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 4880)
  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: pAG61
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        581..924
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1743..1940
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2517..2536)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2518..2536)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2623..2640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2794..3382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3553..4344,4345..4413))
                     /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     4176..4195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4414..4518)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4586..4604
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4642..4664)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4764..4783
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4864..4880,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4864..4880,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgatct gcccgccggg aaggcgaacc cgatcggatg catcctctct
      121 gctgccatga tgctgaagtt gtcgttgaac atggttgctg ccggcgaggc ggtcgagcag
      181 gcagtgcagg aggtgttgga ctcgggagtc agaacgggcg acctgctcgg ctcgagctcc
      241 acttcggagg ttggcgacgc cattgcgctt gcagttaagg aagccttgcg caggcaatcc
      301 gcagctggtc tgagctagcc tcgaggaccc ttctctttag actattctac tcttatgcac
      361 gtaaaaaatt ctaggaaata tgtattaact aggagtaaaa taaccggcta gtggcattca
      421 tatagccgtc tgtttacatc tacatcacac atttcgagtg tatatctcgc aacgttggcg
      481 ttaaataggc agtcaatggc ccgaccattc tatggtgttt aggtcgatgc catctttgta
      541 cgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
      601 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      661 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      721 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
      781 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
      841 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
      901 catcacatcc gaacataaac aaccatgaca gtcaacacta agacctatag tgagagagca
      961 gaaactcatg cctcaccagt agcacagcga ttatttcgat taatggaact gaagaaaacc
     1021 aatttatgtg catcaattga cgttgatacc actaaggaat tccttgaatt aattgataaa
     1081 ttaggtcctt atgtatgctt aatcaagact catattgata taatcaatga tttttcctat
     1141 gaatccacta ttgaaccatt attagaactt tcacgtaaac atcaatttat gatttttgaa
     1201 gatagaaaat ttgctgatat tggtaatacc gtaaagaaac aatatattgg tggagtttat
     1261 aaaattagta gttgggcaga tattaccaat gctcatggtg tcactgggaa tggagtggtt
     1321 gaaggattaa aacagggagc taaagaaacc accaccaacc aagagccaag agggttattg
     1381 atgttagctg aattatcatc agtgggatca ttagcatatg gagaatattc tcaaaaaact
     1441 gttgaaattg ctaaatccga taaggaattt gttattggat ttattgccca acgtgatatg
     1501 ggtggccaag aagaaggatt tgattggctt attatgacac ctggagttgg attagatgat
     1561 aaaggtgatg gattaggaca acaatataga actgttgatg aagttgttag cactggaact
     1621 gatattatca ttgttggtag aggattgttt ggtaaaggaa gagatccaga tattgaaggt
     1681 aaaaggtata gaaatgctgg ttggaatgct tatttgaaaa agactggcca attataatca
     1741 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     1801 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     1861 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     1921 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgagat
     1981 ctgcccgccg ggaaggcgaa cccgatcgga tgcatcctct ctgctgccat gatgctgaag
     2041 ttgtcgttga acatggttgc tgccggcgag gcggtcgagc aggcagtgca ggaggtgttg
     2101 gactcgggag tcagaacggg cgacctgctc ggctcgagct ccacttcgga ggttggcgac
     2161 gccattgcgc ttgcagttaa ggaagccttg cgcaggcaat ccgcagctgg tctgagctag
     2221 cctcgaggac ccttctcttt agactattct actcttatgc acgtaaaaaa ttctaggaaa
     2281 tatgtattaa ctaggagtaa aataaccggc tagtggcatt catatagccg tctgtttaca
     2341 tctacatcac acatttcgag tgtatatctc gcaacgttgg cgttaaatag gcagtcaatg
     2401 gcccgaccat tctatggtgt ttaggtcgat gccatctttg tacgtttaaa cgagctcgaa
     2461 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct
     2521 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg
     2581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4741 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4801 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     4861 acgatttagg tgacactata
//
