LOCUS       sequence-8862-ex        4141 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 4141)
  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 4141)
  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-TRP1
FEATURES             Location/Qualifiers
     source          1..4141
                     /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"
     promoter        802..949
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             950..1624
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             1698..1706
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1778..1797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1779..1797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1884..1901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2814..3605,3606..3674))
                     /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     3437..3456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3675..3779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3847..3865
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3903..3925)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4025..4044
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4125..4141,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4125..4141,1)
                     /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 tgtacaatct tgatccggag cttttctttt tttgccgatt
      841 aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact
      901 attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat
      961 taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc
     1021 cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag
     1081 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
     1141 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
     1201 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
     1261 gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
     1321 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
     1381 tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt
     1441 tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga
     1501 aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt
     1561 ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa
     1621 ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tgcggtgtga
     1681 aataccgcac agatgcgtaa ggaggtttaa acgagctcga attcatcgat gatatcagat
     1741 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     1801 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1861 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2881 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2941 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3001 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3061 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3781 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3841 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3901 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3961 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4021 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
     4081 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     4141 a
//
