LOCUS       sequence-332143-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-yeVenus 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-yeVenus pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    197..221
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             427..1143
                     /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
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    complement(1152..1176)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1238..1257)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1239..1257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1405..1592
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1460..1484)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1604..1621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1604..1620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1613..1635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2056..3402
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3513..4316)
                     /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(4317..4532)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4557..4575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4643..4747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4748..4816,4817..5608)
                     /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(4966..4985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5779..6367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6268..6287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6521..6538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6655..6676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6691..6696,6697..6714,6715..6721)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6729..6745
                     /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     6734..6756
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6753..6769
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6753..6769
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6799..7254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6936..6957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7148..7167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caatagtgaa aatctttttt tcttctatat ctacaaaaac tttttttttc tatcaacctc
       61 gttgataaat tttttcttta acaatcgtta ataattaatt aattggaaaa taaccatttt
      121 ttctctcttt tatacacaca ttcaaaagaa agaaaaaaaa tataccccag cctcgactag
      181 tgaattcgcg gccatcacaa gtttgtacaa aaaagcaggc tgctagcgct accggactca
      241 gatctggtac cccttgcagc ttccagttgt caggaagaga aggctcccat ggatcctgtg
      301 gtggtgctgg ggctctgtct ctcctgtttg cttctccttt cactctggaa acagagctat
      361 gggggaggga agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc
      421 gccaccatgt ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta
      481 gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact
      541 tacggtaaat tgaccttaaa attgatttgt actactggta aattgccagt tccatggcca
      601 accttagtca ctactttagg ttatggtttg caatgttttg ctagataccc agatcatatg
      661 aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt
      721 tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc
      781 ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt
      841 cacaaattgg aatacaacta taactctcac aatgtttaca tcactgctga caaacaaaag
      901 aatggtatca aagctaactt caaaattaga cacaacattg aagatggtgg tgttcaatta
      961 gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac
     1021 cattacttat cctatcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg
     1081 gtcttgttag aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa
     1141 taagcggccg cacccagctt tcttgtacaa agtggtgtag ttaattcatg atggccgctg
     1201 caggtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc
     1261 gcggatccag ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc
     1321 ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga
     1381 tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     1441 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     1501 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     1561 gaggtcgctc ttattgacca cacctctacc ggcatgccaa ttcactggcc gtcgttttac
     1621 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1681 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1741 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     1801 tttcacaccg cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac
     1861 ctgggagttt tccctgaaac agatagtata tttgaacctg tataataata tatagtctag
     1921 cgctttacgg aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca
     1981 taggtaatct tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa
     2041 tagcatatct ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2101 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2161 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2221 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2281 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2341 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2401 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2461 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2521 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2581 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     2641 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     2701 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     2761 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     2821 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     2881 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     2941 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     3001 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa
     3061 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca
     3121 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca
     3181 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat
     3241 gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg
     3301 tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt
     3361 agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt
     3421 caaacatgag aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     3481 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     3541 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     3601 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     3661 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     3721 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     3781 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     3841 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     3901 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     3961 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     4021 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     4081 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     4141 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     4201 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     4261 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     4321 tatcttcgtt tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     4381 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     4441 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa
     4501 aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct
     4561 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4921 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5881 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6541 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6601 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6661 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6721 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6781 caagcttacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6841 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6901 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6961 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7021 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7081 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7141 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7201 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     7261 gccaagcttc ctgaaacgga gaaacataaa caggcattgc tgggatcacc catacatcac
     7321 tctgttttgc ctgacctttt ccggtaattt gaaaacaaac ccggtctcga agcggagatc
     7381 cggcgataat taccgcagaa ataaacccat acacgagacg tagaaccagc cgcacatggc
     7441 cggagaaact cctgcgagaa tttcgtaaac tcgcgcgcat tgcatctgta tttcctaatg
     7501 cggcacttcc aggcctcgat cgagaccgtt tatccattgc ttttttgttg tctttttccc
     7561 tcgttcacag aaagtctgaa gaagctatag tagaactatg agcttttttt gtttctgttt
     7621 tccttttttt tttttttacc tctgtggaaa ttgttactct cacactcttt agttcgtttg
     7681 tttgttttgt ttattccaat tatgaccggt gacgaaacgt ggtcgatggt gggtaccgct
     7741 tatgctcccc tccattagtt tcgattatat aaaaaggcca aatattgtat tattttcaaa
     7801 tgtcctatca ttatcgtcta acatctaatt tctcttaaat tttttctctt tctttcctat
     7861 aacac
//
