LOCUS       sequence-326337-        4514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a yeast codon optimized version of msGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4514)
  AUTHORS   Valbuena FM, Fizgerald I, Strack RL, Andruska N, Smith L, Glick BS
  TITLE     A photostable monomeric superfolder GFP.
  JOURNAL   Traffic. 2020 May 15. doi: 10.1111/tra.12737.
  PUBMED    32415747
REFERENCE   2  (bases 1 to 4514)
  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: "Traffic. 
            2020 May 15. doi: 10.1111/tra.12737."
COMMENT     Sequence Label: yo-msGFP2
FEATURES             Location/Qualifiers
     source          1..4514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             458..1165
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(1194..1211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1194..1210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1203..1225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1419..1438)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1538..1560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1678..2481)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKESRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2482..2697)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2704..2722)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2790..2894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2895..2963,2964..3755)
                     /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     complement(3113..3132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3926..4514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4415..4434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
      421 gcatgcctgc aggtcgactc tagaggatcc aactagtatg gatagcactg agagcttgtt
      481 cacaggtgtt gttcctatct tggtcgaatt agacggtgac gtaaacggtc ataagttcag
      541 tgttagaggt gaaggtgaag gtgacgctac taacggtaaa ttgacattga agtttatttg
      601 tactacaggt aaattgccag ttccttggcc aactttggtt actacattga catacggtgt
      661 tcaatgtttt tcaagatacc cagatcatat gaagcaacat gatttcttta agtctgctat
      721 gccagaaggt tacgttcaag aaagaactat cacttttaaa gatgatggta cttacaagac
      781 aagagcagag gttaagttcg aaggtgacac tttggttaac agaatcgaat tgaagggtat
      841 cgattttaaa gaagatggta atattttggg tcataaattg gaatacaact acaactcaca
      901 taacgtttac attacagctg ataaacaaaa gaatggtatt aaagcaaact tcaagatcag
      961 acataatgtt gaagatggtt ctgttcaatt agcagatcat taccaacaaa acactccaat
     1021 tggtgacggt ccagttttgt taccagataa ccattacttg tctacacaat caaaattgtc
     1081 taaagatcca aacgaaaaga gagatcatat ggttttgtta gaatttgtta ctgctgctgg
     1141 tatcacaggt ggcagtggcg gttctgctag ctaagcggcc gcagctcgaa ttcactggcc
     1201 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1261 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1321 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     1381 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     1441 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     1501 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     1561 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcg ggtaataact gatataatta
     1621 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     1681 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     1741 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     1801 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     1861 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     1921 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     1981 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     2041 aaggcctcta gattcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     2101 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     2161 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     2221 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     2281 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     2341 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     2401 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     2461 cttatatgta gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag
     2521 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     2581 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     2641 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
     2701 ctggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2881 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2941 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3001 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3061 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3121 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3181 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3241 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3301 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3361 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3421 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3481 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3541 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3601 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3661 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3721 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3781 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3841 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3901 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3961 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4021 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4081 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4141 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4201 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4261 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4321 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4381 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4441 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4501 ggagcctatg gaaa
//
