LOCUS       sequence-73685-e        7818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7818)
  TITLE     Series-12:  S cerevisiae Expression Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7818)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Gal1/10 His6-MBP TEV Trp S. cerevisiae express...
FEATURES             Location/Qualifiers
     source          1..7818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(118..186,187..978)
                     /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(336..355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1149..1737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1638..1657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1891..1908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2061..2066,2067..2084,2085..2091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2099..2115
                     /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     2104..2126
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2123..2139
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2123..2139
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2158..2178
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2160..2178
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2215..2231
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2215..2231
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        2518..3182
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2734..2851
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3146..3169
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3216..3233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3243..4343
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4317..4340
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4398..4418
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     4991..5012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5252..5926)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5927..6207)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6224..6243)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6343..6365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6403..6421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      6462..7804
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    6794..6841
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
       61 ccgctcatga gacaataacc ctgataaatg cttcaataaa attgaaaaag gaagagtatg
      121 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      181 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      241 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      301 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      361 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      421 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      481 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      541 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      601 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      661 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      721 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      781 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      841 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      901 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      961 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1021 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1081 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1141 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1201 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1261 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1321 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1381 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1441 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1501 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1561 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1621 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1681 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1741 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1801 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1861 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1921 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1981 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttacctc
     2041 actcattagg caccccaggc tttacacttt atgcttccgg ctcctatgtt gtgtggaatt
     2101 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     2161 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga
     2221 actagtggat cttttgaaag atgatactct ttattgcagc cagttatata agatcttttg
     2281 aaagatgata ctctttattg catgccagtt atataagatc ccccgggctg caggaattcg
     2341 atatcaagct tatcgatacc gtcgaccaat tcgacaggtt atcagcaaca acacagtcat
     2401 atccattctc aattagctct accacagtgt gtgaaccaat gtatccagca ccacctgtaa
     2461 ccaaaacaat tttagaagta ctttcacttt gtaactgagc tgtcatttat attgaatttt
     2521 caaaaattct tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg
     2581 cattaccacc atatacatat ccatatacat atccatatct aatcttactt atatgttgtg
     2641 gaaatgtaaa gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta
     2701 atacgcttaa ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac
     2761 agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa
     2821 acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt
     2881 tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga
     2941 atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta
     3001 attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca
     3061 taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta
     3121 ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa
     3181 accccggatc cgtaaccatg tcaactggtt cttctcacca tcaccatcac catggttctt
     3241 ctatgaaaat cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
     3301 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
     3361 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
     3421 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
     3481 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
     3541 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
     3601 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
     3661 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
     3721 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
     3781 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
     3841 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
     3901 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
     3961 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
     4021 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt
     4081 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     4141 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     4201 ccactatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     4261 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     4321 aagccctgaa agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca
     4381 acaacctcgg gatcgaggaa aacctgtact tccaatccaa tattggaagt ggataactct
     4441 cgaggatcgt aatgactaga ttatataatc agaaatagcc tttggtgatc acttccattt
     4501 tagtcagcat aatagaaagt ttaattacgc gcttttttac attccgacaa agatctgccc
     4561 gagaaaattc ttcgagaaga caacaacaac aaaaaaaata aagcctgggc gaaaaacatc
     4621 gtgcgtttac ataatatctt gaaaagcaaa cagatatata aaccgctatt gaaattacct
     4681 ttttaagtat tcgcctagaa gccttgtttc aagtagcatc aacaacaggt gttaggttaa
     4741 actgctttga ctttttgttc tttaacgaat atcattaaaa aaaaaggaca tctataaaga
     4801 acgcgtaatt cctgcagccc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4861 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4921 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgaacgtt ggagtccacg
     4981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5101 taacaaaaat ttaacgcgaa ttttaacaaa ttattaacgt ttacaatttc ctgatgcggt
     5161 attttctcct tacgcatctg tgcggtattt cacaccgcat agatcggcaa gtgcacaaac
     5221 aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg
     5281 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     5341 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     5401 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     5461 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     5521 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     5581 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     5641 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
     5701 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
     5761 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
     5821 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
     5881 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc caagctgcct
     5941 ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct ccctcttggc
     6001 cctctccttt tcttttttcg accgaattaa ttcttaatcg gcaaaaaaag aaaagctccg
     6061 gatcaagatt gtacgtaagg tgacaagcta tttttcaata aagaatatct tccactactg
     6121 ccatctggcg tcataactgc aaagtacaca tatattacga tgctgtctat taaatgcttc
     6181 ctatattata tatatagtaa tgtcgtttat ggtgcactct cagtacaatc tgctctgatg
     6241 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6301 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6361 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     6421 ttttataggt taatgtcatg ataataatgg tttcttagta tgatccaata tcaaaggaaa
     6481 tgatagcatt gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa
     6541 agggtagtgc tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg
     6601 tactagacta cctttcatcc tacataaata gacgcatata agtacgcatt taagcataaa
     6661 cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg
     6721 cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt
     6781 tcggaaacgc tttgaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
     6841 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
     6901 accggactgt aacgagctac taaaataatg cgaataccgc ttccacaaac attgctcaaa
     6961 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
     7021 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
     7081 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
     7141 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
     7201 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
     7261 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
     7321 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
     7381 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
     7441 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
     7501 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
     7561 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     7621 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
     7681 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
     7741 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
     7801 cgttcaggtg gcactttt
//
