LOCUS       sequence-453902-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymiRFP680 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7543)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7543)
  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 ymiRFP680
FEATURES             Location/Qualifiers
     source          1..7543
                     /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      1601..1848
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1617..1635)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1866..1885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1867..1885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1895..1912)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1895..1911)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1904..1926)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2052..2507
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2139..2158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2349..2370
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2808..3215
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3216..4310
                     /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(4804..4823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4923..4945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4983..5001)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5043..5546
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5583..5687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5688..5756,5757..6548)
                     /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(5906..5925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6719..7307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7208..7227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7461..7478
                     /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 agaatggctg aaggttctgt
      661 tgctagacaa ccagatttgt tgacttgtga tgatgaacca attcatattc caggtgctat
      721 tcaaccacat ggtttgttat tagctttggc tgctgatatg accattgttg ctggttctga
      781 taacttgcct gaattgactg gtttagctat tggtgccttg atcggtagat ctgctgctga
      841 tgttttcgat tctgaaaccc ataatagatt gactattgct ttggctgaac caggtgctgc
      901 tgtcggtgct ccaattaccg tcggtttcac catgcgtaag gatgccggtt tcattggttc
      961 ttggcataga catgatcaat taattttctt ggaattggaa ccaccacaac gtgacgttgc
     1021 cgaaccacag gctttcttcc gtagaaccaa ctctgctatt agaagattac aagctgctga
     1081 aactttggaa tctgcttgtg ctgctgctgc tcaagaagtt cgtaagatta ctggtttcga
     1141 tagagttatg atctacagat ttgcttctga tttctctggt gaagttattg ctgaagatag
     1201 atgtgctgaa gttgaatcta agttgggttt gcattaccca gcttctactg ttccagctca
     1261 agctagaaga ttgtacacta ttaatccagt tagaattatt ccagatatca actacagacc
     1321 agtcccagtt accccagatt tgaacccagt tactggtaga cctattgatt tatctttcgc
     1381 tattttgaga tctgtttccc catgtcattt ggaattcatg agaaacatcg gtatgcatgg
     1441 taccatgtct atttctattt tgagaggtga aagattgtgg ggtttgattg tttgtcatca
     1501 tagaactcca tactacgttg acttggatgg tagacaagcc tgtaagagag ttgctgaaag
     1561 attggctacc caaattggtg ttatggaaga ataactcgag tcatgtaatt agttatgtca
     1621 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     1681 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     1741 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     1801 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca
     1861 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     1921 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1981 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     2041 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2101 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2161 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2221 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2281 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2341 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2401 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2461 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     2521 tttctcctta cgcatctgtg cggtatttca caccgcatat cgaccggtcg aggagaactt
     2581 ctagtatatc tacataccta atattattgc cttattaaaa atggaatccc aacaattaca
     2641 tcaaaatcca cattctcttc aaaatcaatt gtcctgtact tccttgttca tgtgtgttca
     2701 aaaacgttat atttatagga taattatact ctatttctca acaagtaatt ggttgtttgg
     2761 ccgagcggtc taaggcgcct gattcaagaa atatcttgac cgcagttaac tgtgggaata
     2821 ctcaggtatc gtaagatgca agagttcgaa tctcttagca accattattt ttttcctcaa
     2881 cataacgaga acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt
     2941 ttgttaattt cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa
     3001 aaggaaaggt gagagcgccg gaaccggctt ttcatataga atagagaagc gttcatgact
     3061 aaatgcttgc atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc
     3121 caataggtgg ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa
     3181 tatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg
     3241 ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag
     3301 ctatttctga tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg
     3361 ctgctatcga tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg
     3421 ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac
     3481 ctgaacaagg tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat
     3541 gtaactttgc atccgactct cttttagact tatctccaat caagccacaa tttgctaaag
     3601 gtactgactt cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg
     3661 aagacgatgg tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa
     3721 gaatcacaag aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt
     3781 ccttggataa agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa
     3841 ccatcaagaa cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca
     3901 tgatcctagt taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg
     3961 gtgatatcat ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg
     4021 cgtccttggc ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg
     4081 gttctgctcc agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg
     4141 caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag
     4201 ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca
     4261 ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc
     4321 tttttttatg atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga
     4381 aattacaaaa tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat
     4441 ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat tgatactaat
     4501 ggctcaacgt gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg
     4561 caccacacaa aaagttaggt gtaacagaaa atcatgaaac tatgattcct aatttatata
     4621 ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga
     4681 ccatttgatg gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc
     4741 cctcaagaat tttactctgt cagaaacggc cttaacgacg tagtcgatat ggtgcactct
     4801 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4861 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4921 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4981 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga
     5041 cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg
     5101 aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa
     5161 gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa
     5221 caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa atagatatac
     5281 attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac
     5341 agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta
     5401 gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt
     5461 ctttaatttc tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa
     5521 ttataattat ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg
     5581 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5641 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5701 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5761 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5821 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5881 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5941 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6001 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6061 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6121 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6181 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6241 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6301 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6361 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6421 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6481 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6541 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6601 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6661 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6721 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6781 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6841 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6901 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6961 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7021 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7081 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7141 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7201 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7261 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7321 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7381 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7441 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7501 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
