LOCUS       sequence-12733-e        4423 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 4423)
  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 4423)
  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-BIO-kanMX6
FEATURES             Location/Qualifiers
     source          1..4423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      338..525
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(393..417)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            600..1956
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        600..943
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             944..1753
                     /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(1011..1030)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1759..1956
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2060..2079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2061..2079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2166..2183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2337..2925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2417..2436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3096..3887,3888..3956))
                     /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     3719..3738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3957..4061)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4129..4147
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4185..4207)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4307..4326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4407..4423,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4407..4423,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgctggaaa ggccggtgaa ggtgctggaa aggccggtga aggtgaaatc cctgcccctc
      121 ttgctggtac cgtttctaag atactggtaa aagaaggtga cactgttaaa gctggtcaaa
      181 cagttctggt gctggaggct atgaaaatgg agacagaaat taacgctcct actgacggaa
      241 aagttgaaaa ggtgttagtt aaggaaagag atgctgttca aggtggtcaa ggtctaatca
      301 agatcggcgt ttaatgaggc gcgccacttc taaataagcg aatttcttat gatttatgat
      361 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
      421 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
      481 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
      541 taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg
      601 acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat
      661 gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac
      721 attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc
      781 agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa
      841 atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct
      901 tgctaggata cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac
      961 tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1021 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     1081 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1141 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1201 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca
     1261 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1321 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1381 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1441 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     1501 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     1561 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     1621 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     1681 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     1741 tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     1801 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     1861 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     1921 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     1981 catccagttt aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg
     2041 gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     2101 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2161 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2221 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2281 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2341 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2401 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2461 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2521 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2581 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2641 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2701 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2761 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2821 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2881 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2941 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3001 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3061 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3121 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3181 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3241 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3301 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3361 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3421 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3481 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3541 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3601 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3661 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3721 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3781 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3841 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3901 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3961 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4021 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4081 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4141 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4201 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4261 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4321 attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac
     4381 ataaccttat gtatcataca catacgattt aggtgacact ata
//
