LOCUS       sequence-83377-e        5177 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 5177)
  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 5177)
  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: pAG34
FEATURES             Location/Qualifiers
     source          1..5177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            572..2147
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        572..915
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             916..1944
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             916..1944
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1950..2147
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2814..2833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2815..2833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2920..2937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3091..3679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3171..3190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3850..4641,4642..4710))
                     /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     4473..4492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4711..4815)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4883..4901
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4939..4961)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5061..5080
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5161..5177,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5161..5177,1)
                     /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 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt
      961 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     1021 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     1081 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     1141 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt
     1201 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     1261 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     1321 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     1381 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     1441 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     1501 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     1561 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     1621 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     1681 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     1741 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     1801 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     1861 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     1921 actcgtccga gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga
     1981 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2041 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2101 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     2161 actaacgccg ccatccagtg tcgactaggg ttgctgccat cggcctcgct cgcgtctttg
     2221 ccggatagca agagcgcctt tggcctctac gagccctgcc acggctctgc gcccgatctg
     2281 cccgccggga aggcgaaccc gatcggatgc atcctctctg ctgccatgat gctgaagttg
     2341 tcgttgaaca tggttgctgc cggcgaggcg gtcgagcagg cagtgcagga ggtgttggac
     2401 tcgggagtca gaacgggcga cctgctcggc tcgagctcca cttcggaggt tggcgacgcc
     2461 attgcgcttg cagttaagga agccttgcgc aggcaatccg cagctggtct gagctagcct
     2521 cgaggaccct tctctttaga ctattctact cttatgcacg taaaaaattc taggaaatat
     2581 gtattaacta ggagtaaaat aaccggctag tggcattcat atagccgtct gtttacatct
     2641 acatcacaca tttcgagtgt atatctcgca acgttggcgt taaataggca gtcaatggcc
     2701 cgaccattct atggtgttta ggtcgatgcc atctttgtac gtttaaacga gctcgaattc
     2761 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     2821 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     2881 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2941 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3001 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4861 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4921 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4981 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5041 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
     5101 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     5161 atttaggtga cactata
//
