LOCUS       sequence-460515-        7657 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expressing ymScarletI3 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDRF1 ymScarletI3
FEATURES             Location/Qualifiers
     source          1..7657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1547..1571)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1633..1652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1634..1652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1800..1987
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1855..1879)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1999..2016)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1999..2015)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2008..2030)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2451..3797
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3908..4711)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4712..4927)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4952..4970)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5038..5142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5143..5211,5212..6003)
                     /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(5361..5380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6174..6762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6663..6682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6916..6933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7050..7071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7086..7091,7092..7109,7110..7116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7124..7140
                     /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     7129..7151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7148..7164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7148..7164
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7194..7649)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7331..7352)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7543..7562
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc ccatggtcta
      841 gaatggattc tactgaagct gttattaagg aattcatgag atttaaggtt catatggaag
      901 gttctatgaa tggtcatgaa tttgaaattg aaggtgaagg tgaaggtaga ccatatgaag
      961 gtactcaaac tgctaagttg aaggtcacta agggtggtcc attgccattc tcttgggata
     1021 ttttgtctcc acaattcatg tacggttcta gagcttttat taagcatcca gctgacattc
     1081 cagattactg gaagcaatct ttcccagaag gttttaaatg ggaaagagtt atgatctttg
     1141 aagatggtgg tactgtttct gttactcaag atacttcttt ggaagatggt actttgattt
     1201 acaaggttaa gttgagaggt ggtaacttcc caccagatgg tccagttatg caaaagagaa
     1261 ctatgggttg ggaagcctct actgaaagat tgtacccaga agatgttgtt ttaaagggtg
     1321 atattaagat ggctttgaga ttgaaggatg gtggtagata cttggctgat tttaagacta
     1381 cttataaggc taagaagcca gttcaaatgc caggtgcttt caacattgat agaaagttgg
     1441 atattacttc tcataacgaa gattacactg ttgttgaaca atacgaaaga tctgttgcta
     1501 gacattctac tggtggttct ggtggttctt aactcgaggc ggccgcaccc agctttcttg
     1561 tacaaagtgg tgtagttaat tcatgatggc cgctgcaggt cgacctcgag ggggggcccg
     1621 gtacccaatt cgccctatag tgagtcgtat tacgcgcgga tccagctttg gacttcttcg
     1681 ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt
     1741 acgaaaagat ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag
     1801 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     1861 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     1921 tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct
     1981 ctaccggcat gccaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2041 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2101 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     2161 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa tcggatcgta
     2221 cttgttaccc atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata
     2281 gtatatttga acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta
     2341 tttcggttcc tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc
     2401 cggttcattt tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat
     2461 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     2521 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     2581 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2641 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     2701 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     2761 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     2821 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     2881 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     2941 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     3001 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     3061 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     3121 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     3181 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     3241 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     3301 caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt
     3361 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     3421 ctatactttc tagctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa
     3481 acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca
     3541 cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt
     3601 tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc
     3661 tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt
     3721 agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt
     3781 tcctttgata ttggatcgat ccgatgataa gctgtcaaac atgagaattg ggtaataact
     3841 gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata
     3901 cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt
     3961 aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa
     4021 taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt
     4081 ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc
     4141 ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa
     4201 tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat gacaattctg
     4261 ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc
     4321 taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc
     4381 tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt
     4441 cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct
     4501 ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta
     4561 atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt
     4621 ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag
     4681 cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg catgtttttg
     4741 ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc
     4801 gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta
     4861 ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg
     4921 aattgaaaag ctaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4981 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5161 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5281 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5341 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5521 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5701 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5761 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5821 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6001 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6841 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6901 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6961 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7021 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7081 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7141 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttaccgcatc aggaaattgt
     7201 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7261 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7321 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7381 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7441 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7501 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7561 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7621 cgcgcttaat gcgccgctac agggcgcgtc cattcgc
//
