LOCUS       sequence-73680-e        8488 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 8488)
  TITLE     Series-12:  S cerevisiae Expression Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8488)
  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            "Unpublished"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 TEV Ade S. cerevisiae expression ...
FEATURES             Location/Qualifiers
     source          1..8488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(285..305)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(285..303)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(324..340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(324..340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(337..359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    348..364
                     /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)."
     promoter        complement(join(372..378,379..396,397..402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    417..438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(555..572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(726..1314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(806..825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1485..2276,2277..2345))
                     /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     2108..2127
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2346..2450)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2476..3818
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3439..3486)
                     /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)."
     primer_bind     3859..3877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3915..3937)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             4273..5988
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(6193..6648)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6330..6351)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6542..6561
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6774..6796
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6788..6805
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6789..6805
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6815..6834
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6815..6833
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7020..7684
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    7236..7353
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7648..7671
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             7718..7735
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7745..7765
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 aatggacgat atcccgcaag aggccccggc agtaccggca taaccaagcc tatgcctaca
       61 gccttccagg tgacggtgcc gagattgacc atgaacgcct tgttagattt ctacaccggt
      121 gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt tttctttcca
      181 attttttttt tttcgtcatt ataaaaatca ttacgaccga gattcccggg ggaaccacta
      241 gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
      301 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      361 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      421 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      481 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      541 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      601 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      661 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      721 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      781 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      841 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      901 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      961 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1021 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1081 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1141 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     1201 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1261 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1321 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1381 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1441 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1501 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1561 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1621 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1681 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1741 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1801 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1861 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1921 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1981 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2041 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2101 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2161 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2221 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2281 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2341 ctcatactct tcctttttca ttattattga agcatttatc agggttattg tctcatgagc
     2401 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2461 cgaaaagtgc cacctaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2521 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2581 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2641 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2701 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2761 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2821 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2881 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2941 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3001 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3061 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3121 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3181 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3241 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3301 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc attattttag
     3361 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     3421 ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact tcggaatagg
     3481 aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     3541 cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     3601 aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag
     3661 gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
     3721 cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
     3781 tcatccttca atgctatcat ttcctttgat attggatcat actaagaaac cattattatc
     3841 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     3901 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3961 gcggatgccg ggagcagaga attttgtaac accaacataa cactgacatc tttaacaact
     4021 tttaattatg atacatttct tacgtcatga ttgattatta cagctatgct gacaaatgac
     4081 tcttgttgca tggctacgaa ccgggtaata ctaagtgatt gactcttgct gaccttttat
     4141 taagaactaa atggactata ttatggagca tttcatgtat aaattggtgc gtaaaatcgt
     4201 tggatctctc ttctaagtac atcctactat aacaatcaag aaaaacaaga aaatcggaca
     4261 aaacaatcaa gtatggattc tagaacagtt ggtatattag gagggggaca attgggacgt
     4321 atgattgttg aggcagcaaa caggctcaac attaagacgg taatactaga tgctgaaaat
     4381 tctcctgcca aacaaataag caactccaat gaccacgtta atggctcctt ttccaatcct
     4441 cttgatatcg aaaaactagc tgaaaaatgt gatgtgctaa cgattgagat tgagcatgtt
     4501 gatgttccta cactaaagaa tcttcaagta aaacatccca aattaaaaat ttacccttct
     4561 ccagaaacaa tcagattgat acaagacaaa tatattcaaa aagagcattt aatcaaaaat
     4621 ggtatagcag ttacccaaag tgttcctgtg gaacaagcca gtgagacgtc cctattgaat
     4681 gttggaagag atttgggttt tccattcgtc ttgaagtcga ggactttggc atacgatgga
     4741 agaggtaact tcgttgtaaa gaataaggaa atgattccgg aagctttgga agtactgaag
     4801 gatcgtcctt tgtacgccga aaaatgggca ccatttacta aagaattagc agtcatgatt
     4861 gtgaggtctg ttaacggttt agtgttttct tacccaattg tagagactat ccacaaggac
     4921 aatatctgtg acttatgtta tgcgcctgct agagttccgg actccgttca acttaaggcg
     4981 aagttgttgg cagaaaatgc aatcaaatct tttcccggtt gtggtatatt tggtgtggaa
     5041 atgttctatt tagaaacagg ggaattgctt attaacgaaa ttgccccaag gcctcacaac
     5101 tctggacatt ataccattga tgcttgcgtc acttctcaat ttgaagctca tttgagatca
     5161 attttggatt tgccaatgcc aaagaatttc acatctttct ccaccattac aacgaacgcc
     5221 attatgctaa atgttcttgg agacaaacat acaaaagata aagagctaga aacttgcgaa
     5281 agagcattgg cgactccagg ttcctcagtg tacttatatg gaaaagagtc tagacctaac
     5341 agaaaagtag gtcacataaa tatcattgcc tccagtatgg cggaatgtga acaaaggctg
     5401 aactacatta caggtagaac tgatattcca atcaaaatct ctgtcgctca aaagttggac
     5461 ttggaagcaa tggtcaaacc attggttgga atcatcatgg gatcagactc tgacttgccg
     5521 gtaatgtctg ccgcatgtgc ggttttaaaa gattttggcg ttccatttga agtgacaata
     5581 gtctctgctc atagaactcc acataggatg tcagcatatg ctatttccgc aagcaagcgt
     5641 ggaattaaaa caattatcgc tggagctggt ggggctgctc acttgccagg tatggtggct
     5701 gcaatgacac cacttcctgt catcggtgtg cccgtaaaag gttcttgtct agatggagta
     5761 gattctttac attcaattgt gcaaatgcct agaggtgttc cagtagctac cgtcgctatt
     5821 aataatagta cgaacgctgc gctgttggct gtcagactgc ttggcgctta tgattcaagt
     5881 tatacaacga aaatggaaca gtttttatta aagcaagaag aagaagttct tgtcaaagca
     5941 caaaagttag aaactgtcgg ttacgaagct tatctagaaa acaagtaata tataagttta
     6001 ttgatatact tgtacagcaa ataattataa aatgatatac ctatttttta ggctttgtta
     6061 tgattacatc aaatgtggac ttcatacata gaaatcaacg cttacaggtg tcctttttta
     6121 agaatttcat acataagatc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6181 taccgcatca ggaaattgta agcgttaatc ttttgttaaa attcgcgtta aatttttgtt
     6241 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6301 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6361 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6421 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6481 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6541 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6601 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     6661 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6721 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6781 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     6841 gggtaccggg cctcccctgg aggtcgacca attcgacagg ttatcagcaa caacacagtc
     6901 atatccattc tcaattagct ctaccacagt gtgtgaacca atgtatccag caccacctgt
     6961 aaccaaaaca attttagaag tactttcact ttgtaactga gctgtcattt atattgaatt
     7021 ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat
     7081 ggcattacca ccatatacat atccatatac atatccatat ctaatcttac ttatatgttg
     7141 tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag
     7201 taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggtg
     7261 acagccctcc gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
     7321 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
     7381 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac
     7441 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
     7501 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg
     7561 cataaccact ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg
     7621 taataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
     7681 aaaccccgga tccgtaacca tgtcaactgg ttcttctcac catcaccatc accatggttc
     7741 ttctgaaaac ctgtacttcc aatccaatat tggaagtgga taactctcga ggatcgtaat
     7801 gactagatta tataatcaga aatagccttt ggtgatcact tccattttag tcagcataat
     7861 agaaagttta attacgcgct tttttacatt ccgacaaaga tctgcccgag aaaattcttc
     7921 gagaagacaa caacaacaaa aaaaataaag cctgggcgaa aaacatcgtg cgtttacata
     7981 atatcttgaa aagcaaacag atatataaac cgctattgaa attacctttt taagtattcg
     8041 cctagaagcc ttgtttcaag tagcatcaac aacaggtgtt aggttaaact gctttgactt
     8101 tttgttcttt aacgaatatc attaaaaaaa aaggacatct ataaagaatg tctccaaact
     8161 cgtcgaaaac tcgaacggac cagataagct ccatgcctgg tattaatgaa gccacgaagg
     8221 tggaaagtaa gaatgttgtg aaggatgctg ttccaataaa atctgaagtt gaaaccaatg
     8281 gtacatcaat agtaagagaa aaacaagatc cacaggacgg gtgtggtcgc catgatcgcg
     8341 tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg
     8401 gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc
     8461 acgccatagt gactggtgat gctgtcgg
//
