LOCUS       sequence-8861-ex        4664 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 4664)
  AUTHORS   Vorvis C, Markus SM, Lee WL
  TITLE     Photoactivatable GFP tagging cassettes for protein-tracking studies 
            in the budding yeast Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2008 Sep;25(9):651-9.
  PUBMED    18727145
REFERENCE   2  (bases 1 to 4664)
  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: "2008-09"; volume: "25(9)"; pages: "651-9"
COMMENT     Sequence Label: pFA6a-PA-GFP-kanMX6
FEATURES             Location/Qualifiers
     source          1..4664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(76..78,79..81,82..792)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(121..142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(382..401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     729..750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     gene            841..2197
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        841..1184
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1185..1994
                     /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(1252..1271)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2000..2197
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2301..2320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2302..2320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2407..2424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2578..3166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2658..2677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3337..4128,4129..4197))
                     /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     3960..3979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4198..4302)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4370..4388
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4426..4448)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4548..4567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4648..4664,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4648..4664,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      121 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      181 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      241 gtgccctggc ccaccctcgt gaccaccttc agctacggcg tgcagtgctt cagccgctac
      301 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      361 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      421 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      481 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      541 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
      601 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      661 ctgcccgaca accactacct gagccaccag tccgccctga gcaaagaccc caacgagaag
      721 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
      781 gagctgtaca agtagagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
      841 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
      901 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      961 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
     1021 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
     1081 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
     1141 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
     1201 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1261 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     1321 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1381 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1441 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
     1501 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1561 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1621 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1681 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     1741 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1801 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1861 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1921 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1981 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     2041 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     2101 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     2161 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     2221 ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     2281 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     2341 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2521 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2581 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2641 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2701 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2761 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2821 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2881 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2941 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3001 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3061 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3241 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3301 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4321 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4381 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4441 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4501 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4561 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     4621 cataacctta tgtatcatac acatacgatt taggtgacac tata
//
