LOCUS       sequence-73684-e        6663 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 6663)
  TITLE     Series-12:  S cerevisiae Expression Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6663)
  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            "Unpublished"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 Trp S. cerevisiae expression ...
FEATURES             Location/Qualifiers
     source          1..6663
                     /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..3263
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     3835..3856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4096..4770)
                     /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(4771..5052)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5069..5088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5188..5210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5248..5266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5307..6649
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5639..5686
                     /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 ctgaaaacct gtacttccaa tccaatattg gaagtggata actctcgagg atcgtaatga
     3301 ctagattata taatcagaaa tagcctttgg tgatcacttc cattttagtc agcataatag
     3361 aaagtttaat tacgcgcttt tttacattcc gacaaagatc tgcccgagaa aattcttcga
     3421 gaagacaaca acaacaaaaa aaataaagcc tgggcgaaaa acatcgtgcg tttacataat
     3481 atcttgaaaa gcaaacagat atataaaccg ctattgaaat taccttttta agtattcgcc
     3541 tagaagcctt gtttcaagta gcatcaacaa caggtgttag gttaaactgc tttgactttt
     3601 tgttctttaa cgaatatcat taaaaaaaaa ggacatctat aaagaacgcg taattcctgc
     3661 agcccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattttaa caaattatta acgtttacaa tttcctgatg cggtattttc tccttacgca
     4021 tctgtgcggt atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata
     4081 ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc
     4141 ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct
     4201 aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc
     4261 ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt
     4321 cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact
     4381 gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa
     4441 ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc
     4501 attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt
     4561 cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac
     4621 cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga
     4681 atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc
     4741 agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac
     4801 gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt
     4861 ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt
     4921 aaggtgacaa gctatttttc aataaagaat atcttccact actgcacatc tggcgtcata
     4981 actgcaaagt acacatatat tacgatgctg tctattaaat gcttcctata ttatatatat
     5041 agtaatgtcg tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5101 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5161 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5221 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5281 tcatgataat aatggtttct tagtatgatc caatatcaaa ggaaatgata gcattgaagg
     5341 atgagactaa tccaattgag gagtggcagc atatagaaca gctaaagggt agtgctgaag
     5401 gaagcatacg ataccccgca tggaatggga taatatcaca ggaggtacta gactaccttt
     5461 catcctacat aaatagacgc atataagtac gcatttaagc ataaacacgc actatgccgt
     5521 tcttctcatg tatatatata tacaggcaac acgcagatat aggtgcgacg tgaacagtga
     5581 gctgtatgtg cgcagctcgc gttgcatttt cggaagcgct cgttttcgga aacgctttga
     5641 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcagag cgcttttgaa
     5701 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     5761 gctactaaaa taatgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     5821 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     5881 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     5941 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     6001 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     6061 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     6121 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     6181 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     6241 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     6301 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     6361 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     6421 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     6481 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
     6541 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     6601 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgttc aggtggcact
     6661 ttt
//
