LOCUS       sequence-453900-        7525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing yemiRFP670 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7525)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7525)
  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 yemiRFP670
FEATURES             Location/Qualifiers
     source          1..7525
                     /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      1583..1830
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1599..1617)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1848..1867)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1849..1867)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1877..1894)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1877..1893)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1886..1908)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2034..2489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2121..2140)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2331..2352
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2790..3197
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3198..4292
                     /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(4786..4805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4905..4927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4965..4983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5025..5528
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5565..5669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5670..5738,5739..6530)
                     /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(5888..5907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6701..7289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7190..7209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7443..7460
                     /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 acatgaagaa attcatttgg ctggttctat
      721 tcaaccacat ggtgctttgt tggtcgtttc tgaacatgac catagagtta ttcaagcctc
      781 tgctaatgcc gctgaattct taaacttggg ttctgtttta ggtgttccat tagctgaaat
      841 tgatggtgat ttgttaatta agattttacc acatttagat ccaaccgctg aaggtatgcc
      901 agttgctgtt agatgtagaa tcggtaatcc ttccactgaa tactgtggtt tgatgcatag
      961 accaccagaa ggtggtttga ttattgaatt ggaaagagct ggtccatcta ttgatttatc
     1021 cggtactttg gctccagctt tggaaagaat tagaactgct ggttctttaa gagctttgtg
     1081 tgatgacact gttttgttat tccaacaatg tactggttat gatagagtta tggtttacag
     1141 attcgatgaa caaggtcatg gtttagtttt ctctgaatgt catgtccctg gtttggaatc
     1201 ttacttcggt aatagatacc catcctctac tgttccacaa atggctagac aattatacgt
     1261 tagacaaaga gttagagttt tggtcgatgt tacttaccaa ccagttccat tggaaccaag
     1321 attatcccca ttgactggta gagatttaga tatgtctggt tgtttcttaa gatctatgtc
     1381 tccttgtcat ttgcaattct tgaaggatat gggtgttaga gctactttgg ccgtttcttt
     1441 agttgttggt ggtaagttgt ggggtttggt tgtttgtcac cattacttgc caagattcat
     1501 tagattcgaa ttaagagcta tctgtaagag attggctgaa agaattgcta ctagaattac
     1561 cgctttagaa tcttaactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1621 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1681 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1741 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     1801 gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc tatagtgagt
     1861 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1921 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     1981 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     2041 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2101 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2161 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2221 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2281 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2341 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2401 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2461 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     2521 tgcggtattt cacaccgcat atcgaccggt cgaggagaac ttctagtata tctacatacc
     2581 taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct
     2641 tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag
     2701 gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc
     2761 ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg
     2821 caagagttcg aatctcttag caaccattat ttttttcctc aacataacga gaacacacag
     2881 gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc
     2941 gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagcgc
     3001 cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat
     3061 acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat
     3121 cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata tatatatttc
     3181 aaggatatac cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg
     3241 ttggtcaaga aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt
     3301 ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag
     3361 gtgttccact tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag
     3421 gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa
     3481 aaatccgtaa agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact
     3541 ctcttttaga cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg
     3601 tcagagaatt agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg
     3661 tcgcttggga tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg
     3721 ctttcatggc cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg
     3781 ttttggcctc ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgaattcc
     3841 ctacattgaa ggttcaacat caattgattg attctgccgc catgatccta gttaagaacc
     3901 caacccacct aaatggtatt ataatcacca gcaacatgtt tggtgatatc atctccgatg
     3961 aagcctccgt tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc
     4021 cagacaagaa caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc
     4081 caaagaataa ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt
     4141 cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg
     4201 caggtatcag aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg
     4261 tcgccgaaga agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt
     4321 acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat
     4381 gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa
     4441 aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga
     4501 aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag
     4561 gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa
     4621 aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa
     4681 gtcaatacct tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct
     4741 gtcagaaacg gccttaacga cgtagtcgat atggtgcact ctcagtacaa tctgctctga
     4801 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4861 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4921 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4981 atttttatag gttaatgtca tgataataat ggtttcttag gacggatcgc ttgcctgtaa
     5041 cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat
     5101 ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg
     5161 gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca
     5221 tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt
     5281 aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt
     5341 cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc
     5401 cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta
     5461 ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt atttttatag
     5521 cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5581 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5641 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5701 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5761 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5821 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5881 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5941 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6001 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6061 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6121 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6181 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6241 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6301 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6361 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6421 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6481 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6541 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6601 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6661 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6721 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6781 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6841 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6901 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6961 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7021 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7081 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7141 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7201 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7261 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7321 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7381 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7441 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7501 cgcgttggcc gattcattaa tgcag
//
