LOCUS       sequence-127134-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagging with mTFP1 using URA3 as a selection marker in S.
            cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  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 4713)
  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: POM42-mTFP1
FEATURES             Location/Qualifiers
     source          1..4713
                     /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    1538..1571
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1610..2314
                     /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(2350..2369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2351..2369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2456..2473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2627..3215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2707..2726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3386..4177,4178..4246))
                     /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     4009..4028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4247..4351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4419..4437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4475..4497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4597..4616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4697..4713,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4697..4713,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc ccaatacaac agatcacgtg
      121 atctttttgt aagatgaagt tgaagtagtg ttgcaccgtg ccaatgcagg tggctattag
      181 attaaatatg tgatttgttc tattaagttt cctgtataat taatggggag cgctgattct
      241 cttttggtac gcttcccatc cagcatttct gtatctttca ccttcaacct taggatctct
      301 acccttggcg aaaagtcctc tgccaacaat gatgatatct gatccaccac ttacaacttc
      361 gtcgacggtt ctgtactgct gacccaatgc atcgcctttg tcgtctaaac ctacacctgg
      421 ggtcatgatt agccaatcaa acccttcttc tcttcctccc atatcgttct gagcaatgaa
      481 cccaataacg aaatctttat cactctttgc aatatcaacg gtacccttag tatattcacc
      541 gtgtgctaga gaacccttgg aagacaattc agcaagcatc aataatcccc ttggttcttt
      601 ggtgacctct tgcgcacctt gtttcaagcc agcaacaata ccagcaccag taaccccgtg
      661 ggcgttggtg atatcagacc attctgcgat acggtaaacg cccgatgtat attgtaattt
      721 gactgtgtta ccgatatcgg cgaattttct gtcctcaaat atcaagaact tgtatttctc
      781 tgccaatgct ttcaatggaa cgacagtacc ctcataactg aaatcatcca agatatcaac
      841 gtgtgttttc aaaaggcaaa tgtatggacc caacgtttca acaagtttca atagctcatc
      901 agtcgaacga acgtcaagag aagcacacaa attggtcttc ttttcatcca ttaaacgtaa
      961 aagtttcgat gcaaccggac ttgcatgagt ctcagctcta ctggtatatg attttgtgga
     1021 catggtgcaa ctaattgacg ggagtgtatt gacgctggcg tactggcttt cacaaaatgg
     1081 cccaatcaca accacatctt agatagttga aatgacttta gataacatca attgagatga
     1141 gcttaatcat gtcaaagcta aaagtgtcac catgaacgac aattcttaag caaatcacgt
     1201 gatatagatc cacgaataac caccatttga tgctcgaggc aagtaatgtg tgtaaaaaaa
     1261 tgcgttacca ccatccaatg cagaccgatc ttctacccag aatcacatat atttatgtac
     1321 cgagtacctt ttttctatct tccaattgct tctcccatat gattgtctcc gtaagctcga
     1381 aatttctaag ttggatttta atcttcacgc aggatgacag ttcgatgagc ttctgaggag
     1441 tgtttagaac ataatcattt atccatggtc tatctcttct tgtcgctttt tctcctcgat
     1501 agaacctaaa taaaacgagc tctcgagaac ccttaatata acttcgtata atgtatgcta
     1561 tacgaagtta ttaggtgata tcatgcgtac gctgcaggtc gacggatccg tgagcaaggg
     1621 cgaggagacc acaatgggcg taatcaagcc cgacatgaag atcaagctga agatggaggg
     1681 caacgtgaat ggccacgcct tcgtgatcga gggcgagggc gagggcaagc cctacgacgg
     1741 caccaacacc atcaacctgg aggtgaagga gggagccccc ctgcccttct cctacgacat
     1801 tctgaccacc gcgttcgcct acggcaacag ggccttcacc aagtaccccg acgacatccc
     1861 caactacttc aagcagtcct tccccgaggg ctactcttgg gagcgcacca tgaccttcga
     1921 ggacaagggc atcgtgaagg tgaagtccga catctccatg gaggaggact ccttcatcta
     1981 cgagatacac ctcaagggcg agaacttccc ccccaacggc cccgtgatgc agaagaagac
     2041 caccggctgg gacgcctcca ccgagaggat gtacgtgcgc gacggcgtgc tgaagggcga
     2101 cgtcaagcac aagctgctgc tggagggcgg cggccaccac cgcgttgact tcaagaccat
     2161 ctacagggcc aagaaggcgg tgaagctgcc cgactatcac tttgtggacc accgcatcga
     2221 gatcctgaac cacgacaagg actacaacaa ggtgaccgtt tacgagagcg ccgtggcccg
     2281 caactccacc gacggcatgg acgagctgta taagactagt ggcctatgcg gccgcggatc
     2341 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     2401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2821 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3061 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4381 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4441 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4501 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4561 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4621 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     4681 gtatcataca catacgattt aggtgacact ata
//
