LOCUS       sequence-469817-        7290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymNeongreen in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7290)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7290)
  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 ymNeongreen
FEATURES             Location/Qualifiers
     source          1..7290
                     /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"
     regulatory      886..895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1348..1595
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1364..1382)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1613..1632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1614..1632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1642..1659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1642..1658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1651..1673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1799..2254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1886..1905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2096..2117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2555..2962
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2963..4057
                     /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(4551..4570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4670..4692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4730..4748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4790..5293
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5330..5434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5435..5503,5504..6295)
                     /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(5653..5672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6466..7054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6955..6974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7208..7225
                     /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 atggtctcta agggtgaaga agataacatg
      661 gcttctttgc cagccaccca cgagttgcac atcttcggtt ccattaacgg tgtcgacttc
      721 gatatggtcg gtcaaggtac cggtaaccca aacgacggtt acgaagaatt gaacttgaag
      781 tctaccaagg gcgacttgca gttctcccct tggattttgg ttccacacat cggttacggt
      841 ttccaccaat acttgccata cccagacggt atgtccccat tccaagccgc catggttgac
      901 ggttctggtt accaagtcca cagaaccatg caattcgaag acggtgcttc cttgaccgtg
      961 aactacagat acacctacga aggttctcac atcaagggcg aagcccaagt taagggtacc
     1021 ggtttccctg ctgacggtcc agtcatgact aactccttga ccgctgctga ctggtgtcgt
     1081 tccaagaaga cctacccaaa cgataagacc atcatttcca ctttcaagtg gtcctacacc
     1141 accggtaacg gcaagagata ccgctctacc gctcgtacca cctacacctt cgccaagcca
     1201 atggctgcca actacttgaa gaaccagcct atgtacgtct tccgtaagac cgagctgaag
     1261 cattccaaga ctgaattgaa cttcaaggaa tggcaaaagg ctttcaccga cgtcatgggt
     1321 atggacgaat tgtacaagta actcgagtca tgtaattagt tatgtcacgc ttacattcac
     1381 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1441 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     1501 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     1561 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag
     1621 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     1801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1981 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2221 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     2281 atctgtgcgg tatttcacac cgcatatcga ccggtcgagg agaacttcta gtatatctac
     2341 atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat
     2401 tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt
     2461 tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa
     2521 ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta
     2581 agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca
     2641 cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag
     2701 aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag
     2761 agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc
     2821 acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta
     2881 gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat
     2941 atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt tgccaggtga
     3001 ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt
     3061 tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc
     3121 tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg atgccgtttt
     3181 gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt
     3241 actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc
     3301 cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt
     3361 tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga
     3421 tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat
     3481 ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct tggataaagc
     3541 taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga
     3601 attccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa
     3661 gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc
     3721 cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc
     3781 tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga
     3841 tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa tgatgttgaa
     3901 attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt
     3961 ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga
     4021 tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata
     4081 tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg
     4141 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
     4201 gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
     4261 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
     4321 gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc
     4381 tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag
     4441 tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt
     4501 actctgtcag aaacggcctt aacgacgtag tcgatatggt gcactctcag tacaatctgc
     4561 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4621 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4681 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4741 cgcctatttt tataggttaa tgtcatgata ataatggttt cttaggacgg atcgcttgcc
     4801 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
     4861 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
     4921 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
     4981 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
     5041 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
     5101 caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg
     5161 atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt
     5221 ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt
     5281 tatagcacgt gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5941 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7081 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7141 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7201 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7261 ccccgcgcgt tggccgattc attaatgcag
//
