LOCUS       sequence-452187-        7282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymScarletI3 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7282)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7282)
  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 ymScarletI3
FEATURES             Location/Qualifiers
     source          1..7282
                     /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      1340..1587
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1356..1374)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1605..1624)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1606..1624)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1634..1651)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1634..1650)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1643..1665)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1791..2246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1878..1897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2088..2109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2547..2954
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2955..4049
                     /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(4543..4562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4662..4684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4722..4740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4782..5285
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5322..5426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5427..5495,5496..6287)
                     /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(5645..5664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6458..7046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6947..6966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7200..7217
                     /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 accatggtct agaatggatt ctactgaagc
      661 tgttattaag gaattcatga gatttaaggt tcatatggaa ggttctatga atggtcatga
      721 atttgaaatt gaaggtgaag gtgaaggtag accatatgaa ggtactcaaa ctgctaagtt
      781 gaaggtcact aagggtggtc cattgccatt ctcttgggat attttgtctc cacaattcat
      841 gtacggttct agagctttta ttaagcatcc agctgacatt ccagattact ggaagcaatc
      901 tttcccagaa ggttttaaat gggaaagagt tatgatcttt gaagatggtg gtactgtttc
      961 tgttactcaa gatacttctt tggaagatgg tactttgatt tacaaggtta agttgagagg
     1021 tggtaacttc ccaccagatg gtccagttat gcaaaagaga actatgggtt gggaagcctc
     1081 tactgaaaga ttgtacccag aagatgttgt tttaaagggt gatattaaga tggctttgag
     1141 attgaaggat ggtggtagat acttggctga ttttaagact acttataagg ctaagaagcc
     1201 agttcaaatg ccaggtgctt tcaacattga tagaaagttg gatattactt ctcataacga
     1261 agattacact gttgttgaac aatacgaaag atctgttgct agacattcta ctggtggttc
     1321 tggtggttct taactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1381 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1441 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1501 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1561 ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt
     1621 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1681 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1741 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     1801 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1861 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1921 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1981 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2041 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2101 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2161 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2221 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     2281 ggtatttcac accgcatatc gaccggtcga ggagaacttc tagtatatct acatacctaa
     2341 tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac attctcttca
     2401 aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata tttataggat
     2461 aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct aaggcgcctg
     2521 attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg taagatgcaa
     2581 gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg
     2641 cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc
     2701 tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg
     2761 aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact
     2821 tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt
     2881 cttactttct aacttttctt accttttaca tttcagcaat atatatatat atatttcaag
     2941 gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg
     3001 gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca
     3061 atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg
     3121 ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg
     3181 ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa
     3241 tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca tccgactctc
     3301 ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca
     3361 gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg
     3421 cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga atggccgctt
     3481 tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatgttt
     3541 tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac gaattcccta
     3601 cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa
     3661 cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag
     3721 cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag
     3781 acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa
     3841 agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat
     3901 tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag
     3961 gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg
     4021 ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca
     4081 taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt
     4141 catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg
     4201 aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa
     4261 agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg
     4321 taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt tctctaaaaa
     4381 aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc
     4441 aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc
     4501 agaaacggcc ttaacgacgt agtcgatatg gtgcactctc agtacaatct gctctgatgc
     4561 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4621 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4681 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4741 tttataggtt aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt
     4801 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
     4861 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
     4921 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
     4981 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
     5041 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
     5101 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
     5161 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
     5221 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
     5281 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5401 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5461 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5521 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5581 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5641 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5701 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5761 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5821 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5881 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5941 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6001 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6061 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6121 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6181 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6241 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6301 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6361 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6421 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6481 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6541 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6601 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6661 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6721 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6781 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6841 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6901 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6961 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7021 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7081 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7141 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7201 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7261 gttggccgat tcattaatgc ag
//
