LOCUS       sequence-453899-        7299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymTurquoise2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7299)
  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"
COMMENT     Sequence Label: P415 ymTurquoise2
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    52..73
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(88..93,94..111,112..118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    126..142
                     /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     131..153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     150..166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     150..166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     185..205
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        187..205
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        223..621
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      1357..1604
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1373..1391)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1622..1641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1623..1641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1651..1668)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1651..1667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1660..1682)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1808..2263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1895..1914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2105..2126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2564..2971
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2972..4066
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4560..4579)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4679..4701
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4739..4757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4799..5302
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5339..5443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5444..5512,5513..6304)
                     /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(5662..5681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6475..7063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6964..6983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7217..7234
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
       61 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      181 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctcatagctt caaaatgttt
      241 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
      301 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
      361 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
      421 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt
      481 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
      541 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
      601 agcatagcaa tctaatctaa gttttctaga atggttagta aaggtgaaga attgttcact
      661 ggtgtagtgc caattttagt agaattagat ggtgacgtta acggtcataa gttctctgta
      721 agcggtgaag gtgaaggtga cgcaacatac ggtaaattaa ctttgaaatt catttgtact
      781 acaggtaaat taccagttcc atggccaact ttggttacta ctttgtcttg gggtgttcaa
      841 tgtttcgcta gatacccaga tcatatgaag caacatgatt tctttaagtc tgctatgcca
      901 gaaggttacg ttcaagaaag aactatcttc ttcaaggatg atggtaacta taagactaga
      961 gctgaagtta agttcgaagg agatacttta gttaatagaa ttgaattgaa gggtattgat
     1021 tttaaggaag atggtaatat tttgggtcat aagttggaat ataattactt ctctgataac
     1081 gtttacatta ctgctgataa gcaaaagaac ggtattaagg cgaacttcaa gattagacat
     1141 aacattgaag atggtggtgt tcaattagct gatcattacc aacaaaacac tccaattggt
     1201 gacggtccag ttttgttgcc agataaccat tacttgtcta ctcaatctaa gttgtctaag
     1261 gatccaaacg aaaagagaga tcatatggtt ttgttggaat tcgttactgc tgctgggatc
     1321 actttaggta tggatgaatt gtataaataa ctcgagtcat gtaattagtt atgtcacgct
     1381 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     1441 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     1501 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     1561 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     1621 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     1681 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1741 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1801 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1861 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1921 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1981 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2041 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2101 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2161 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2221 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     2281 tccttacgca tctgtgcggt atttcacacc gcatatcgac cggtcgagga gaacttctag
     2341 tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa
     2401 aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa
     2461 cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga
     2521 gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca
     2581 ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata
     2641 acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt
     2701 taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg
     2761 aaaggtgaga gcgccggaac cggcttttca tatagaatag agaagcgttc atgactaaat
     2821 gcttgcatca caatacttga agttgacaat attatttaag gacctattgt tttttccaat
     2881 aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata
     2941 tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt
     3001 gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat
     3061 ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc
     3121 tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga
     3181 tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga
     3241 acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa
     3301 ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac
     3361 tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga
     3421 cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat
     3481 cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt
     3541 ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat
     3601 caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat
     3661 cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga
     3721 tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc
     3781 cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc
     3841 tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat
     3901 gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa
     3961 aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga
     4021 agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt
     4081 tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt
     4141 acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac atacatttat
     4201 caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct
     4261 caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc
     4321 acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt tatatattgg
     4381 aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg acctgaccat
     4441 ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga aagttccctc
     4501 aagaatttta ctctgtcaga aacggcctta acgacgtagt cgatatggtg cactctcagt
     4561 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4621 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4681 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4741 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttaggacgga
     4801 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     4861 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     4921 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     4981 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     5041 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     5101 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     5161 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt
     5221 aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat
     5281 aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg
     5341 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5401 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5461 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5521 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5581 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5641 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5701 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5761 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5821 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5881 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5941 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6001 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6061 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6121 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6181 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6241 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6301 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6361 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7141 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7201 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7261 accgcctctc cccgcgcgtt ggccgattca ttaatgcag
//
