LOCUS       sequence-491306-        7685 bp    DNA     circular SYN 31-OCT-2025
DEFINITION  Expresses mTq2 in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7685)
  TITLE     mTurquoise2 pDRF1-GW
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7685)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 31, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: mTurquoise2 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     280..301
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      343..352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(369..393)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      1201..1656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1288..1307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1498..1519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1686..1702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1686..1702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1699..1721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1710..1726
                     /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)."
     promoter        complement(join(1734..1740,1741..1758,1759..1764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1779..1800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1917..1934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2088..2676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2168..2187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2847..3638,3639..3707))
                     /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     3470..3489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3708..3812)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3880..3898
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3923..4138
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4139..4942
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      5053..6399
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6820..6842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6834..6851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6835..6851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(6863..7050)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6971..6995
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     7198..7217
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7198..7216
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7279..7303
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(7312..7685,1..340,341..343,344..346))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7357..7378)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
       61 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
      121 ggtagcgggc gaagcactgc acgccccagg acagggtggt cacgagggtg ggccagggca
      181 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
      241 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
      301 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta
      361 gcgctagcag cctgcttttt tgtacaaact tgtgatggcc gcgaattcac tagtcgaggc
      421 tggggtatat tttttttctt tcttttgaat gtgtgtataa aagagagaaa aaatggttat
      481 tttccaatta attaattatt aacgattgtt aaagaaaaaa tttatcaacg aggttgatag
      541 aaaaaaaaag tttttgtaga tatagaagaa aaaaagattt tcactattgg tgttatagga
      601 aagaaagaga aaaaatttaa gagaaattag atgttagacg ataatgatag gacatttgaa
      661 aataatacaa tatttggcct ttttatataa tcgaaactaa tggaggggag cataagcggt
      721 acccaccatc gaccacgttt cgtcaccggt cataattgga ataaacaaaa caaacaaacg
      781 aactaaagag tgtgagagta acaatttcca cagaggtaaa aaaaaaaaaa aggaaaacag
      841 aaacaaaaaa agctcatagt tctactatag cttcttcaga ctttctgtga acgagggaaa
      901 aagacaacaa aaaagcaatg gataaacggt ctcgatcgag gcctggaagt gccgcattag
      961 gaaatacaga tgcaatgcgc gcgagtttac gaaattctcg caggagtttc tccggccatg
     1021 tgcggctggt tctacgtctc gtgtatgggt ttatttctgc ggtaattatc gccggatctc
     1081 cgcttcgaga ccgggtttgt tttcaaatta ccggaaaagg tcaggcaaaa cagagtgatg
     1141 tatgggtgat cccagcaatg cctgtttatg tttctccgtt tcaggaagct tggcgaatgg
     1201 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1261 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1321 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1381 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1441 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1501 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1561 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1621 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtaag cttggcgtaa
     1681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2521 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3841 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3901 tttcgtcttc aagaattagc ttttcaattc atcatttttt ttttattctt ttttttgatt
     3961 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     4021 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
     4081 cccagtattc ttaacccaac tgcacagaac aaaaacatgc aggaaacgaa gataaatcat
     4141 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
     4201 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
     4261 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
     4321 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
     4381 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
     4441 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     4501 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     4561 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     4621 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     4681 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     4741 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     4801 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     4861 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     4921 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     4981 acaaattaga gcttcaattt aattatatca gttattaccc aattctcatg tttgacagct
     5041 tatcatcgga tcgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca
     5101 attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac
     5161 cccgcatgga atgggataat atcacaggag gtactagact acctttcatc ctacataaat
     5221 agacgcatat aagtacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata
     5281 tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca
     5341 gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga
     5401 agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc
     5461 gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa
     5521 tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg
     5581 tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact
     5641 cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt
     5701 aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta
     5761 gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca
     5821 acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg
     5881 ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga
     5941 agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac
     6001 cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga
     6061 gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt
     6121 agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct
     6181 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     6241 tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca
     6301 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     6361 gcatttttgt tctacaaaat gaagcacaga tgcttcgtta acaaagatat gctattgaag
     6421 tgcaagatgg aaacgcagaa aatgaaccgg ggatgcgacg tgcaagatta cctatgcaat
     6481 agatgcaata gtttctccag gaaccgaaat acatacattg tcttccgtaa agcgctagac
     6541 tatatattat tatacaggtt caaatatact atctgtttca gggaaaactc ccaggttcgg
     6601 atgttcaaaa ttcaatgatg ggtaacaagt acgatccgat tatgcggtgt gaaataccgc
     6661 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     6721 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     6781 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     6841 cgacggccag tgaattggca tgccggtaga ggtgtggtca ataagagcga cctcatgcta
     6901 tacctgagaa agcaacctga cctacaggaa agagttactc aagaataaga attttcgttt
     6961 taaaacctaa gagtcacttt aaaatttgta tacacttatt ttttttataa cttatttaat
     7021 aataaaaatc ataaatcata agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc
     7081 aacgatttga cccttttcca tcttttcgta aatttctggc aaggtagaca agccgacaac
     7141 cttgattgga gacttgacca aacctctggc gaagaagtcc aaagctggat ccgcgcgtaa
     7201 tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga cctgcagcgg
     7261 ccatcatgaa ttaactacac cactttgtac aagaaagctg ggtgcggccg cttacttgta
     7321 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     7381 atcgcgcttc tcgttggggt ctttgctcag cttggactgg gtgctcaggt agtggttgtc
     7441 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     7501 cacgccgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg
     7561 cttgtcggcg gtgatataga cgttgtcgct aaagtagttg tactccagct tgtgccccag
     7621 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     7681 gccct
//
