LOCUS       sequence-9656-ex        4429 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 4429)
  AUTHORS   Funakoshi M, Hochstrasser M
  TITLE     Small epitope-linker modules for PCR-based C-terminal tagging in 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2009 Feb 25. 26(3):185-192.
  PUBMED    19243080
REFERENCE   2  (bases 1 to 4429)
  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. 2009
            Feb 25. 26(3):185-192."
COMMENT     Sequence Label: pFA6a-6xGLY-His-tag-hphMX4
FEATURES             Location/Qualifiers
     source          1..4429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..96
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      120..307
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(175..199)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            382..1957
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        382..725
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             726..1754
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             726..1754
                     /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      1760..1957
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2064..2083)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2065..2083)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2170..2187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2341..2929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2421..2440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3100..3891,3892..3960))
                     /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     3723..3742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3961..4065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4133..4151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4189..4211)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4311..4330
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4411..4429
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4411..4428
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaca tcatcaccat caccactagg gcgcgccact tctaaataag
      121 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      181 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      241 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
      301 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgccttgtcc
      361 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      421 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      481 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      541 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      601 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      661 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      721 caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
      781 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
      841 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      901 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
      961 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     1021 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     1081 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     1141 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     1201 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     1261 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1321 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1381 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1441 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1501 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1561 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1621 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     1681 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1741 gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     1801 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     1861 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     1921 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     1981 ccatccagtg tcgaaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     2041 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     2101 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2161 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2221 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2281 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2341 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2401 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2461 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2521 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2581 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2641 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2701 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2761 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2821 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2881 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2941 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3001 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3061 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3721 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4021 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4081 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4141 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4201 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4261 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4321 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     4381 atacataacc ttatgtatca tacacatacg atttaggtga cactataga
//
