LOCUS       sequence-47373-e        4787 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 4787)
  AUTHORS   Tang X, Punch JJ, Lee WL
  TITLE     A CAAX motif can compensate for the PH domain of Num1 for cortical 
            dynein attachment.
  JOURNAL   Cell Cycle. 2009 Oct 1;8(19):3182-90. Epub 2009 Oct 4.
  PUBMED    19755860
REFERENCE   2  (bases 1 to 4787)
  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: "Cell 
            Cycle."; date: "2009-10-1"; volume: "8(19)"; pages: "3182-90. Epub 
            2009 Oct 4"
COMMENT     Sequence Label: pKT0128-RAS2 C-term
FEATURES             Location/Qualifiers
     source          1..4787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..718
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      791..978
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(846..870)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1053..2253
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1053..1396
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1397..2050
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2056..2253
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2357..2376)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2358..2376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2463..2480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2714..2733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3393..4184,4185..4253))
                     /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     4016..4035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4254..4358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4426..4444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4482..4504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4604..4623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4704..4722
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4704..4721
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taacatgtct aaaggtgaag aattattcac tggtgttgtc ccaattttgg ttgaattaga
       61 tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg atgctactta
      121 cggtaaattg accttaaaat ttatttgtac tactggtaaa ttgccagttc catggccaac
      181 cttagtcact actttaactt atggtgttca atgtttttct agatacccag atcatatgaa
      241 acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa gaactatttt
      301 tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag gtgatacctt
      361 agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca ttttaggtca
      421 caaattggaa tacaactata actctcacaa tgtttacatc atggctgaca aacaaaagaa
      481 tggtatcaaa gttaacttca aaattagaca caacattgaa gatggttctg ttcaattagc
      541 tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac cagacaacca
      601 ttacttatcc actcaatctg ccttatccaa agatccaaac gaaaagagag accacatggt
      661 cttgttagaa tttgttactg ctgctggtat tacccatggt attgatgaat tgtacaaagg
      721 tgctggtgct ggatcgggtg gctgttgtat tataagttaa gtacaaataa ggcgcgccac
      781 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      841 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      901 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      961 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
     1021 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
     1081 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
     1141 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
     1201 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
     1261 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
     1321 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     1381 ccgaacataa acaaccatgg gtaggagggc ttttgtagaa agaaatacga acgaaacgaa
     1441 aatcagcgtt gccatcgctt tggacaaagc tcccttacct gaagagtcga attttattga
     1501 tgaacttata acttccaagc atgcaaacca aaagggagaa caagtaatcc aagtagacac
     1561 gggaattgga ttcttggatc acatgtatca tgcactggct aaacatgcag gctggagctt
     1621 acgactttac tcaagaggtg atttaatcat cgatgatcat cacactgcag aagatactgc
     1681 tattgcactt ggtattgcat tcaagcaggc tatgggtaac tttgccggcg ttaaaagatt
     1741 tggacatgct tattgtccac ttgacgaagc tctttctaga agcgtagttg acttgtcggg
     1801 acggccctat gctgttatcg atttgggatt aaagcgtgaa aaggttgggg aattgtcctg
     1861 tgaaatgatc cctcacttac tatattcctt ttcggtagca gctggaatta ctttgcatgt
     1921 tacctgctta tatggtagta atgaccatca tcgtgctgaa agcgctttta aatctctggc
     1981 tgttgccatg cgcgcggcta ctagtcttac tggaagttct gaagtcccaa gcacgaaggg
     2041 agtgttgtaa agagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     2101 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     2161 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     2221 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     2281 ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc
     2341 gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg
     2401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2821 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3061 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4381 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4441 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4501 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4561 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4621 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
     4681 accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc gccagctgaa
     4741 gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gtttaat
//
