LOCUS       sequence-12736-e        4237 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 4237)
  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 4237)
  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-RGS18HIS-kanMX6
FEATURES             Location/Qualifiers
     source          1..4237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..101
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             75..104
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             78..107
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             81..110
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             84..113
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             87..116
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             90..119
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             93..122
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             96..125
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      152..339
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(207..231)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            414..1770
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        414..757
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             758..1567
                     /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(825..844)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1573..1770
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1874..1893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1875..1893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1980..1997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2151..2739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2231..2250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2910..3701,3702..3770))
                     /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     3533..3552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3771..3875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3943..3961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3999..4021)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4121..4140
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4221..4237,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4221..4237,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatc atcatcacca ccatcatcat catcaccacc
      121 atcattaatg aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat
      181 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
      241 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
      301 atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag
      361 ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
      421 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
      481 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
      541 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
      601 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
      661 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
      721 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa agactcacgt
      781 ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      841 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
      901 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
      961 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1021 tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     1081 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1141 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1201 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1261 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     1321 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1381 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1441 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1501 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1561 tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     1621 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     1681 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     1741 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     1801 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     1861 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     1921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2341 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2581 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3301 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3901 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3961 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4021 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4081 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4141 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     4201 ttatgtatca tacacatacg atttaggtga cactata
//
