LOCUS       sequence-9668-ex        4447 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4447)
  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 4447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-HSV-hphMX4
FEATURES             Location/Qualifiers
     source          1..4447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..114
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     terminator      138..325
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(193..217)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            400..1975
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        400..743
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             744..1772
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             744..1772
                     /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      1778..1975
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2082..2101)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2083..2101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2188..2205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2359..2947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2439..2458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3118..3909,3910..3978))
                     /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     3741..3760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3979..4083)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4151..4169
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4207..4229)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4329..4348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4429..4447
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4429..4446
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaag ccagccagaa ctcgccccgg aagaccccga ggattagggc
      121 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
      181 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
      241 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
      301 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
      361 gtttagcttg ccttgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
      421 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
      481 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
      541 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
      601 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
      661 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
      721 atcacatccg aacataaaca accatgggta aaaagcctga actcaccgcg acgtctgtcg
      781 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
      841 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
      901 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
      961 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     1021 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     1081 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     1141 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     1201 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     1261 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     1321 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     1381 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     1441 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     1501 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     1561 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     1621 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     1681 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     1741 gccccagcac tcgtccgagg gcaaaggaat aatcagtact gacaataaaa agattcttgt
     1801 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     1861 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     1921 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     1981 gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat cgatgatatc
     2041 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     2101 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2161 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2521 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     2581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2761 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     2821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2881 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2941 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3001 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3061 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3121 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3181 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3241 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3301 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3361 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3421 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3481 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3541 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3601 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3661 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3721 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3781 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3841 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3901 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3961 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4021 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4081 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4141 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4201 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4261 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4321 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
     4381 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     4441 ctataga
//
