LOCUS       sequence-12726-e        4447 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 4447)
  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 4447)
  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-HBH-kanMX6
FEATURES             Location/Qualifiers
     source          1..4447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..89
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..335
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      362..549
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(417..441)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            624..1980
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        624..967
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             968..1777
                     /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(1035..1054)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1783..1980
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2084..2103)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2085..2103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2190..2207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2361..2949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2441..2460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3120..3911,3912..3980))
                     /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     3743..3762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3981..4085)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4153..4171
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4209..4231)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4331..4350
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4431..4447,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4431..4447,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatg ctggaaaggc cggtgaaggt gaaatccctg
      121 cccctcttgc tggtaccgtt tctaagatac tggtaaaaga aggtgacact gttaaagctg
      181 gtcaaacagt tctggtgctg gaggctatga aaatggagac agaaattaac gctcctactg
      241 acggaaaagt tgaaaaggtg ttagttaagg aaagagatgc tgttcaaggt ggtcaaggtc
      301 taatcaagat cggcgttcat catcaccacc atcattaatg aggcgcgcca cttctaaata
      361 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
      421 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
      481 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
      541 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
      601 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
      661 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
      721 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
      781 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
      841 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
      901 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
      961 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     1021 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1081 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1141 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1201 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1261 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1321 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1381 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     1441 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1501 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1561 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1621 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1681 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1741 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     1801 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     1861 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     1921 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     1981 ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata
     2041 tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta
     2101 ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc
     2161 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2221 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2281 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2521 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     2581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2761 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     2821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3421 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3481 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3541 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3601 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3661 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3721 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3781 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3841 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3901 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3961 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4021 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4081 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4141 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4201 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4261 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4321 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg
     4381 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga
     4441 cactata
//
