LOCUS       sequence-8863-ex        4551 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 4551)
  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 4551)
  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-His3MX6
FEATURES             Location/Qualifiers
     source          1..4551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(74..76,77..79,80..790)
                     /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(119..140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(380..399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     727..748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        839..1182
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1183..1881
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1885..2082
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2186..2205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2187..2205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2292..2309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2463..3051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2543..2562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3222..4013,4014..4082))
                     /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     3845..3864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4083..4187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4255..4273
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4311..4333)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4433..4452
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4533..4551
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4533..4550
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaag
       61 gcgcgccaga tccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      181 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      241 gccctggccc accctcgtga ccaccttcag ctacggcgtg cagtgcttca gccgctaccc
      301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      541 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag
      601 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      661 gcccgacaac cactacctga gccaccagtc cgccctgagc aaagacccca acgagaagcg
      721 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
      781 gctgtacaag tagagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga
      841 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
      901 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
      961 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
     1021 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     1081 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     1141 gctaggatac agttctcaca tcacatccga acataaacaa ccatggcaga accagcccaa
     1201 aaaaagcaaa aacaaactgt tcaggagcgc aaggcgttta tctcccgtat cactaatgaa
     1261 actaaaattc aaatcgctat ttcgctgaat ggtggttata ttcaaataaa agattcgatt
     1321 cttcctgcaa agaaggatga cgatgtagct tcccaagcta ctcagtcaca ggtcatcgat
     1381 attcacacag gtgttggctt tttggatcat atgatccatg cgttggcaaa acactctggt
     1441 tggtctctta ttgttgaatg tatcggtgac ctgcacattg acgatcacca tactaccgaa
     1501 gattgcggta tcgcattagg gcaagcgttc aaagaagcaa tgggtgctgt ccgtggtgta
     1561 aaaagattcg gtactgggtt cgcaccattg gatgaggcgc tatcacgtgc cgtagtcgat
     1621 ttatctaata gaccatttgc tgtaatcgac cttggattga agagagagat gattggtgat
     1681 ttatccactg aaatgattcc acactttttg gaaagtttcg cggaggcggc cagaattact
     1741 ttgcatgttg attgtctgag aggtttcaac gatcaccaca gaagtgagag tgcgttcaag
     1801 gctttggctg ttgccataag agaagctatt tctagcaatg gcaccaatga cgttccctca
     1861 accaaaggtg ttttgatgtg aagtactgac aataaaaaga ttcttgtttt caagaacttg
     1921 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     1981 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2041 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2101 cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     2161 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     2221 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2641 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2881 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4201 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4261 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4321 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4381 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4441 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     4501 taatacataa ccttatgtat catacacata cgatttaggt gacactatag a
//
