LOCUS       sequence-47106-e        4873 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 3110-01
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..767
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     terminator      788..975
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(843..867)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1050..2406
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1050..1393
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1394..2203
                     /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(1461..1480)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2209..2406
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2510..2529)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2511..2529)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2616..2633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2787..3375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2867..2886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3546..4337,4338..4406))
                     /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     4169..4188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4407..4511)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4579..4597
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4635..4657)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4757..4776
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4857..4873,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4857..4873,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaacatga gtgcgattaa gccagacatg aagatcaaac
      121 tccgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat ggtacaggca
      181 agccttttga gggaaaacag agtatggatc ttgaagtcaa agagggcgga cctctgcctt
      241 ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc gccaaatatc
      301 cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa
      361 gcttgacttt cgaagacggg ggcatttgca ttgccagaaa cgacataaca atggaagggg
      421 acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta
      481 tgcagaagaa gacgctgaaa tgggagccct ccactgagaa aatgtatgtg cgtgatggag
      541 tgctgacggg tgatattcat atggctttgt tgcttgaagg aaatgcccat taccgatgtg
      601 acttcagaac tacttacaaa gctaaggaga agggtgtcaa gttaccaggc taccactttg
      661 tggaccactg cattgagatt ttaagccatg acaaagatta caacaaggtt aagctgtatg
      721 agcatgctgt tgctcattct ggattgcctg acaatgccag acgatagggc gcgccacttc
      781 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
      841 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
      901 taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga
      961 ccacacctct accggcagat ccgctaggga taacagggta atatagatct gtttagcttg
     1021 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
     1081 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
     1141 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
     1201 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
     1261 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
     1321 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
     1381 aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa
     1441 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1501 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1561 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1621 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1681 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1741 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1801 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1861 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1921 ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     1981 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2041 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2101 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2161 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa
     2221 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     2281 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     2341 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     2401 atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc gaattcatcg
     2461 atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     2521 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     2581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     3001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4501 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4561 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4621 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4681 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4741 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata
     4801 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     4861 aggtgacact ata
//
