LOCUS       sequence-127167-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagging with mTFP1 using LEU2 as a selection marker in S.
            cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  AUTHORS   Higuchi-Sanabria R, Garcia EJ, Tomoiaga D, Munteanu EL, Feinstein P,
            Pon LA
  TITLE     Characterization of Fluorescent Proteins for Three- and Four-Color 
            Live-Cell Imaging in S. cerevisiae.
  JOURNAL   PLoS One. 2016 Jan 4;11(1):e0146120. doi: 
            10.1371/journal.pone.0146120. eCollection 2016.
  PUBMED    26727004
REFERENCE   2  (bases 1 to 5550)
  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: "PLoS One.";
            date: "2016-01-4"; volume: "11(1):e0146120. doi"; pages: " 
            10.1371/journal.pone.0146120. eCollection 2016"
COMMENT     Sequence Label: POM43-mTFP1
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2375..2408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2447..3151
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT
                     NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE
                     DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG
                     DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV
                     ARNSTDGMDELYK"
     primer_bind     complement(3187..3206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3188..3206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3293..3310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3464..4052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3544..3563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4223..5014,5015..5083))
                     /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     4846..4865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5084..5188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5256..5274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5312..5334)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5434..5453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5534..5550,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5534..5550,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc cgcaggctaa ccggaacctg
      121 tattatttag tttatgctac gttaaataaa gacctttcgt tcacataact gaatgtgtaa
      181 tggccttgag atttcaagca taccaagttg gtggagacgg ggtcgttaca aaagactctt
      241 taagccaaga tttccttgac agccttggcg atagcatcgc caacctcagt ggtagagtta
      301 gaaccaccaa ggtcaccggt tctgacacct gcatccaaga catttctaac agcttcttca
      361 agagccctac cttcttcaac caaatccaag gataacttca acatcatagc tgcagataag
      421 atggtagcaa ttgggttaac cttgtttgct ggtaaatctg gggcagaacc atgacatggt
      481 tcgtacaaac cgaatgcctt gttagtgtca ggtagggaag ctagagatgc agaaggtaat
      541 aaacccaaag aacctggaat aacagaggct tcatcggaga taatatcacc aaacatgttg
      601 ttggtaataa caacaccgtt tagcttagtt ggtgatttaa ccaaaatcat agcagcagag
      661 tcgatcaatt ggtgctgaac agttaattgt gggaactcag tcttgatggt ttcttcaaca
      721 gtctttctcc acaatctgga agaggcaagc acgttagcct tgtcaagaga ccagattggt
      781 aatggtgggt tttgttgcaa tgccaagaaa gcagccattc ttgtaattct ttgaacttca
      841 ggaacactgt atttctcaga gtcccaagca actccgtcac cttcatcttc ttttctttca
      901 ccaaagtaga taccaccaac caattctcta acgacgacga aatcggtacc ctttgcatat
      961 tcaggcttca aaggagaaag atctagtaaa gaatcagaag caaagttaca tggtctcaag
     1021 ttggcgtata gacccaattc ctttctgatc ttcaatagac cttgttctgg tctaactgcg
     1081 cccgtacccc attttggacc accaacagca cctagtaaga cagcatcggc tttcttagag
     1141 gcttctagag cttcatctgg taaaggagtg ccagtggcat cgatggcagc acccccgatc
     1201 aagtgatgtt ggaaattgaa cttaatttct ggacggactt cagcaatggc attcaagacc
     1261 ttaatagctt cgtcagtaac ttctttaccg acgtgatcac ccggtaggac aacgatattc
     1321 ttagacatgg tataatctgt gtagtgtggg atacttttta cttcttcaaa taggtatcaa
     1381 caataaaaat aaatcgaagc aaatgtagga atgcgttaaa gcagatgtac ttacttagag
     1441 tacatagata tatttatata attcaatata taaaagtata tgaagcatct gatgttgaac
     1501 ctgtcatgac tctaaactgg aatgggtagt tatgcttctg aaggttttcc gttagaaaat
     1561 gagtgcaggg atcaggcccg gaaccggttt tagcctgaaa ggaaaagaaa aaaatgcggt
     1621 gattactagc acgtgactgc gctgaattgg aatccaccac aagtacagcg aatggcaaga
     1681 gggaagggga atatattgtg cctggcggta gtctgctata tcaaatgtta aacataacta
     1741 aatgcagtag tgaatagagt aatgtatttc tacatatgtg ggtagggcca atgggagccc
     1801 gatgtctatt gtacagatat tttcctttta tagttaatca attcaacttg gaaatcattg
     1861 agtcgatatc tacttggata actggagccg tggcagcgtt tgaaatttga cgagtcttct
     1921 tgttttgttc gcgcattttc agtttattca aacgggagat ggcaccgaat ccactgactt
     1981 ctctgttcaa tttgaatggt aataggaaga aagtacattt cgagaaattt tcgcccaacg
     2041 tcgaaatctc atggatgata tcctcgatac aaatgacacc ttcatcacct aatttctctt
     2101 caacgatatt gttgtcattc aagattatct cttttggttc tttatcttct ggcctttgcc
     2161 acattattct agatctcttt tggattaaag aacggataga tgcaagtgaa ggttgtccga
     2221 taacaatata tggtgataca agtttcaaca atgggtacac gtccttggta agtttgatga
     2281 aaacaccagt gttcaactca acaagtctca aaagagataa cactttgtaa gcctttgctg
     2341 ggatcttcac agcgagctct cgagaaccct taatataact tcgtataatg tatgctatac
     2401 gaagttatta ggtgatatca tgcgtacgct gcaggtcgac ggatccgtga gcaagggcga
     2461 ggagaccaca atgggcgtaa tcaagcccga catgaagatc aagctgaaga tggagggcaa
     2521 cgtgaatggc cacgccttcg tgatcgaggg cgagggcgag ggcaagccct acgacggcac
     2581 caacaccatc aacctggagg tgaaggaggg agcccccctg cccttctcct acgacattct
     2641 gaccaccgcg ttcgcctacg gcaacagggc cttcaccaag taccccgacg acatccccaa
     2701 ctacttcaag cagtccttcc ccgagggcta ctcttgggag cgcaccatga ccttcgagga
     2761 caagggcatc gtgaaggtga agtccgacat ctccatggag gaggactcct tcatctacga
     2821 gatacacctc aagggcgaga acttcccccc caacggcccc gtgatgcaga agaagaccac
     2881 cggctgggac gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga agggcgacgt
     2941 caagcacaag ctgctgctgg agggcggcgg ccaccaccgc gttgacttca agaccatcta
     3001 cagggccaag aaggcggtga agctgcccga ctatcacttt gtggaccacc gcatcgagat
     3061 cctgaaccac gacaaggact acaacaaggt gaccgtttac gagagcgccg tggcccgcaa
     3121 ctccaccgac ggcatggacg agctgtacaa gactagtggc ctatgcggcc gcggatctgc
     3181 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     3241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     3661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5281 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5341 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5401 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5461 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     5521 tcatacacat acgatttagg tgacactata
//
