LOCUS       sequence-9665-ex        4441 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 4441)
  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 4441)
  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-VSV-G-hphMX4
FEATURES             Location/Qualifiers
     source          1..4441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..108
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      132..319
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(187..211)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            394..1969
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        394..737
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             738..1766
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             738..1766
                     /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      1772..1969
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2076..2095)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2077..2095)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2182..2199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2353..2941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2433..2452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3112..3903,3904..3972))
                     /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     3735..3754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3973..4077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4145..4163
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4201..4223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4323..4342
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4423..4441
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4423..4440
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga gcagaaactc atctcagaag aggatctgta gggcgcgcca
      121 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
      181 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
      241 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta
      301 ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag
      361 cttgccttgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
      421 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
      481 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
      541 aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg
      601 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
      661 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
      721 tccgaacata aacaaccatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
      781 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
      841 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
      901 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
      961 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     1021 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     1081 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     1141 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     1201 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     1261 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     1321 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     1381 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     1441 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     1501 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     1561 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     1621 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     1681 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     1741 gcactcgtcc gagggcaaag gaataatcag tactgacaat aaaaagattc ttgttttcaa
     1801 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     1861 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     1921 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg
     1981 atactaacgc cgccatccag tgtcgaaaac gagctcgaat tcatcgatga tatcagatcc
     2041 actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc
     2101 gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2521 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2761 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4081 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4141 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4201 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4261 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4321 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
     4381 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
     4441 a
//
