LOCUS       sequence-47112-e        4215 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 4215)
  AUTHORS   Young CL, Raden DL, Caplan JL, Czymmek KJ, Robinson AS
  TITLE     Cassette series designed for live-cell imaging of proteins and 
            high-resolution techniques in yeast.
  JOURNAL   Yeast. 2012 Mar;29(3-4):119-36. doi: 10.1002/yea.2895. Epub 2012 Apr
            4.
  PUBMED    22473760
REFERENCE   2  (bases 1 to 4215)
  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."; 
            date: "2012-03"; volume: "29(3-4):119-36. doi"; pages: " 
            10.1002/yea.2895. Epub 2012 Apr 4"
COMMENT     Sequence Label: pCY 3140-02
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     gene            170..1745
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        170..513
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             514..1542
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             514..1542
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1548..1745
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1852..1871)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1853..1871)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1958..1975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2129..2717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2209..2228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2888..3679,3680..3748))
                     /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     3511..3530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3749..3853)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3921..3939
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3977..3999)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4099..4118
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4199..4215,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4199..4215,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaactatc catatgatgt tcctgactat gcttaaggcg
      121 cgccagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
      181 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
      241 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
      301 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc
      361 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
      421 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
      481 cagttctcac atcacatccg aacataaaca accatgggta aaaagcctga actcaccgcg
      541 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
      601 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
      661 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
      721 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
      781 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
      841 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
      901 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
      961 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     1021 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     1081 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     1141 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     1201 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     1261 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     1321 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     1381 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     1441 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     1501 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aatcagtact gacaataaaa
     1561 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     1621 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     1681 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     1741 tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat
     1801 cgatgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     1861 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     1921 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     2341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     2581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2761 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2821 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2881 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2941 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3001 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3061 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3901 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3961 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4021 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4081 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca
     4141 tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     4201 ttaggtgaca ctata
//
