LOCUS       sequence-12729-e        4516 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 4516)
  AUTHORS   Tagwerker C, Zhang H, Wang X, Larsen LS, Lathrop RH, Hatfield GW, 
            Auer B, Huang L, Kaiser P
  TITLE     HB tag modules for PCR-based gene tagging and tandem affinity 
            purification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2006 Jun . 23(8):623-32.
  PUBMED    16823883
REFERENCE   2  (bases 1 to 4516)
  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. 2006
            Jun . 23(8):623-32."
COMMENT     Sequence Label: pFA6a-HTB-kanMX6
FEATURES             Location/Qualifiers
     source          1..4516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..89
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             117..137
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      431..618
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(486..510)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            693..2049
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        693..1036
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1037..1846
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1104..1123)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1852..2049
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2153..2172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2154..2172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2259..2276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2430..3018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2510..2529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3189..3980,3981..4049))
                     /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     3812..3831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4050..4154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4222..4240
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4278..4300)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4400..4419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4500..4516,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4500..4516,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatg actacgatat acccacaacc gccagcgaga
      121 atctgtactt tcaatcagag ttaaaaaccg ccgcactagc tcaacatgac gaggctgctg
      181 gaaaggccgg tgaaggtgaa atccctgccc ctcttgctgg taccgtttct aagatactgg
      241 taaaagaagg tgacactgtt aaagctggtc aaacagttct ggtgctggag gctatgaaaa
      301 tggagacaga aattaacgct cctactgacg gaaaagttga aaaggtgtta gttaaggaaa
      361 gagatgctgt tcaaggtggt caaggtctaa tcaagatcgg cgtttaatga ggcgcgccac
      421 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      481 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      541 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      601 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
      661 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      721 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
      781 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
      841 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
      901 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
      961 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     1021 ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc
     1081 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     1141 tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     1201 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     1261 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     1321 caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     1381 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     1441 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     1501 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     1561 agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg
     1621 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     1681 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     1741 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     1801 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat
     1861 aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt
     1921 ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca
     1981 gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc
     2041 gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag ctcgaattca
     2101 tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag
     2161 tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg
     2221 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2281 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2341 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2401 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2461 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2521 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2581 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     2641 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2701 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2761 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2821 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2881 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2941 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3001 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3061 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3121 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3181 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3241 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3301 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3361 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3421 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3481 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3541 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3601 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3661 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3721 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3781 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3841 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3901 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3961 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4021 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4081 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4141 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4201 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4261 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4321 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4381 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac
     4441 atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga
     4501 tttaggtgac actata
//
