LOCUS       sequence-332102-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-mVenus to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7865)
  AUTHORS   Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B
  TITLE     In vivo characterisation of fluorescent proteins in budding yeast.
  JOURNAL   Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z.
  PUBMED    30783202
REFERENCE   2  (bases 1 to 7865)
  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: "Sci Rep."; 
            date: "2019-02-19"; volume: "9(1):2234. doi"; pages: " 
            10.1038/s41598-019-38913-z"
COMMENT     Sequence Label: cyterm-ymVenus pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             452..1168
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    complement(1177..1201)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1263..1282)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1264..1282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1430..1617
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1485..1509)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1629..1646)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1629..1645)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1638..1660)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2081..3427
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3538..4341)
                     /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"
     promoter        complement(4342..4557)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4582..4600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4668..4772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4773..4841,4842..5633)
                     /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(4991..5010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5804..6392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6293..6312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6546..6563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6680..6701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6716..6721,6722..6739,6740..6746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6754..6770
                     /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     6759..6781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6778..6794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6778..6794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6824..7279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6961..6982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7173..7192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttttctctt tctttcctat aacaccaata gtgaaaatct ttttttcttc tatatctaca
       61 aaaacttttt ttttctatca acctcgttga taaatttttt ctttaacaat cgttaataat
      121 taattaattg gaaaataacc attttttctc tcttttatac acacattcaa aagaaagaaa
      181 aaaaatatac cccagcctcg actagtgaat tcgcggccat cacaagtttg tacaaaaaag
      241 caggctgcta gcgctaccgg actcagatct ggtacccctt gcagcttcca gttgtcagga
      301 agagaaggct cccatggatc ctgtggtggt gctggggctc tgtctctcct gtttgcttct
      361 cctttcactc tggaaacaga gctatggggg agggaagctt cgaattctgc agtcgacggt
      421 accgcgggcc cgggatccac cggtcgccac catgtctaaa ggtgaagaat tattcactgg
      481 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
      541 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattga tttgtactac
      601 tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg gtttgcaatg
      661 ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
      721 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
      781 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
      841 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
      901 ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa
      961 cattgaagat ggtggtgttc aattagctga ccattatcaa caaaatactc caattggtga
     1021 tggtccagtc ttgttaccag acaaccatta cttatcctat caatctaaat tatccaaaga
     1081 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     1141 ccatggtatg gatgaattgt acaaataagc ggccgcaccc agctttcttg tacaaagtgg
     1201 tgtagttaat tcatgatggc cgctgcaggt cgacctcgag ggggggcccg gtacccaatt
     1261 cgccctatag tgagtcgtat tacgcgcgga tccagctttg gacttcttcg ccagaggttt
     1321 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
     1381 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     1441 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     1501 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     1561 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     1621 gccaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1681 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1741 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     1801 tttctcctta cgcatctgtg cggtatttca caccgcataa tcggatcgta cttgttaccc
     1861 atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga
     1921 acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc
     1981 tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt
     2041 tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca
     2101 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2161 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2221 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2281 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2341 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     2401 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     2461 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     2521 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     2581 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     2641 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     2701 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     2761 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     2821 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     2881 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     2941 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3001 gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3061 tagctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
     3121 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
     3181 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
     3241 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
     3301 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
     3361 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
     3421 ttggatcgat ccgatgataa gctgtcaaac atgagaattg ggtaataact gatataatta
     3481 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     3541 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     3601 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     3661 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     3721 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     3781 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     3841 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     3901 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     3961 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     4021 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     4081 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     4141 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     4201 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     4261 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     4321 cttatatgta gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag
     4381 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     4441 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     4501 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
     4561 ctaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4621 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4741 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4801 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4861 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4921 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4981 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5041 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5101 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5161 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5221 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5281 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5341 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5401 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5461 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5521 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5581 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5641 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5941 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6541 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     6601 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6661 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6721 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6781 gaaacagcta tgaccatgat tacgccaagc ttaccgcatc aggaaattgt aagcgttaat
     6841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7261 gcgccgctac agggcgcgtc cattcgccaa gcttcctgaa acggagaaac ataaacaggc
     7321 attgctggga tcacccatac atcactctgt tttgcctgac cttttccggt aatttgaaaa
     7381 caaacccggt ctcgaagcgg agatccggcg ataattaccg cagaaataaa cccatacacg
     7441 agacgtagaa ccagccgcac atggccggag aaactcctgc gagaatttcg taaactcgcg
     7501 cgcattgcat ctgtatttcc taatgcggca cttccaggcc tcgatcgaga ccgtttatcc
     7561 attgcttttt tgttgtcttt ttccctcgtt cacagaaagt ctgaagaagc tatagtagaa
     7621 ctatgagctt tttttgtttc tgttttcctt tttttttttt ttacctctgt ggaaattgtt
     7681 actctcacac tctttagttc gtttgtttgt tttgtttatt ccaattatga ccggtgacga
     7741 aacgtggtcg atggtgggta ccgcttatgc tcccctccat tagtttcgat tatataaaaa
     7801 ggccaaatat tgtattattt tcaaatgtcc tatcattatc gtctaacatc taatttctct
     7861 taaat
//
