LOCUS       sequence-9657-ex        4121 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 4121)
  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 4121)
  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-S-tag-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..123
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      147..334
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(202..226)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        409..752
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             753..1451
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1455..1652
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1756..1775)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1757..1775)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1862..1879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2033..2621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2113..2132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2792..3583,3584..3652))
                     /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     3415..3434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3653..3757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3825..3843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3881..3903)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4003..4022
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4103..4121
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4103..4120
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac
      121 agctagggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
      181 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
      241 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
      301 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
      361 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
      421 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
      481 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
      541 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
      601 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
      661 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
      721 agttctcaca tcacatccga acataaacaa ccatggcaga accagcccaa aaaaagcaaa
      781 aacaaactgt tcaggagcgc aaggcgttta tctcccgtat cactaatgaa actaaaattc
      841 aaatcgctat ttcgctgaat ggtggttata ttcaaataaa agattcgatt cttcctgcaa
      901 agaaggatga cgatgtagct tcccaagcta ctcagtcaca ggtcatcgat attcacacag
      961 gtgttggctt tttggatcat atgatccatg cgttggcaaa acactctggt tggtctctta
     1021 ttgttgaatg tatcggtgac ctgcacattg acgatcacca tactaccgaa gattgcggta
     1081 tcgcattagg gcaagcgttc aaagaagcaa tgggtgctgt ccgtggtgta aaaagattcg
     1141 gtactgggtt cgcaccattg gatgaggcgc tatcacgtgc cgtagtcgat ttatctaata
     1201 gaccatttgc tgtaatcgac cttggattga agagagagat gattggtgat ttatccactg
     1261 aaatgattcc acactttttg gaaagtttcg cggaggcggc cagaattact ttgcatgttg
     1321 attgtctgag aggtttcaac gatcaccaca gaagtgagag tgcgttcaag gctttggctg
     1381 ttgccataag agaagctatt tctagcaatg gcaccaatga cgttccctca accaaaggtg
     1441 ttttgatgtg aagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     1501 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     1561 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     1621 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc
     1681 cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg
     1741 cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc
     1801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2221 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2461 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3781 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     3841 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3901 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3961 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4021 tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa
     4081 ccttatgtat catacacata cgatttaggt gacactatag a
//
