LOCUS       sequence-47082-e        4237 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 4237)
  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 4237)
  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 3010-07
FEATURES             Location/Qualifiers
     source          1..4237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..797
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             87..797
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(114..142)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     737..757
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     gene            854..1778
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     promoter        854..1197
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1198..1572
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1581..1778
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1874..1893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1875..1893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1980..1997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2151..2739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2231..2250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2910..3701,3702..3770))
                     /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     3533..3552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3771..3875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3943..3961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3999..4021)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4121..4140
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4221..4237,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4221..4237,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaacagta aaggagaaga acttttcact ggagttgtcc
      121 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
      181 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
      241 tacctgttcc atggccaaca cttgtcacta ctttgacttg gggtgttcaa tgctttgcta
      301 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg
      361 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
      421 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
      481 atggaaacat tcttggacac aaattggaat acaacgctat ttcagataat gtatacatca
      541 ctgcagacaa acaaaagaat ggaatcaaag ctaatttcaa aattagacac aacattgaag
      601 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
      661 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
      721 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acactaggca
      781 tggatgaact atacaaatag ggcgcgccag atctgtttag cttgcctcgt ccccgccggg
      841 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
      901 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
      961 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     1021 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     1081 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     1141 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     1201 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     1261 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     1321 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     1381 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     1441 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     1501 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     1561 gagcaggact gataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     1621 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     1681 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     1741 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     1801 gccatcccga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     1861 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     1921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2341 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2581 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3301 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3901 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3961 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4021 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4081 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4141 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     4201 ttatgtatca tacacatacg atttaggtga cactata
//
