LOCUS       sequence-9651-ex        4109 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 4109)
  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 4109)
  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. 2009
            Feb 25. 26(3):185-192."
COMMENT     Sequence Label: pFA6a-6xGLY-T7-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..111
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     terminator      135..322
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(190..214)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        397..740
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             741..1439
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1443..1640
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1744..1763)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1745..1763)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1850..1867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2021..2609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2101..2120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2780..3571,3572..3640))
                     /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     3403..3422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3641..3745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3813..3831
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3869..3891)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3991..4010
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4091..4109
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4091..4108
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaat ggctagcatg actggtggac agcaaatggg ttagggcgcg
      121 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
      181 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
      241 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
      301 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
      361 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
      421 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
      481 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
      541 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
      601 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
      661 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      721 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc
      781 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt
      841 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg
      901 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt
      961 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta
     1021 tcggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc
     1081 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg
     1141 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctaataga ccatttgctg
     1201 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac
     1261 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag
     1321 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag
     1381 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa
     1441 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     1501 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     1561 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     1621 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
     1681 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
     1741 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     1801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1861 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1921 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1981 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     2221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2461 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2761 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2821 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2881 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2941 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3001 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3061 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3121 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3181 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3241 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3301 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3361 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3421 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3481 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3541 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3601 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3661 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3721 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     3781 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3841 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3901 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3961 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4021 accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca
     4081 tacacatacg atttaggtga cactataga
//
