LOCUS       sequence-8860-ex        4637 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 4637)
  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 4637)
  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: pDH3-PA-GFP
FEATURES             Location/Qualifiers
     source          1..4637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(49..51,52..54,55..765)
                     /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(94..115)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(355..374)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     702..723
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     gene            814..2170
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        814..1157
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1158..1967
                     /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(1225..1244)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1973..2170
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2274..2293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2275..2293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2380..2397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2551..3139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2631..2650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3310..4101,4102..4170))
                     /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     3933..3952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4171..4275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4343..4361
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4399..4421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4521..4540
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4621..4637,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4621..4637,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatccat ggtgagcaag
       61 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      121 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      181 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      241 ttcagctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      301 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      361 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      421 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      481 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcc
      541 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      601 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagccac
      661 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      721 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagag atctgtttag
      781 cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
      841 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
      901 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
      961 aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg
     1021 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
     1081 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
     1141 tccgaacata aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt
     1201 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1261 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1321 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1381 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1441 tcaccactgc gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag
     1501 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1561 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1621 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1681 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     1741 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1801 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1861 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     1921 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa
     1981 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     2041 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     2101 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     2161 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc
     2221 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     2281 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     2341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     2761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4321 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4381 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4441 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4501 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
     4561 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     4621 atttaggtga cactata
//
