LOCUS       sequence-46743-e        6881 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 6881)
  AUTHORS   Blount BA, Weenink T, Vasylechko S, Ellis T
  TITLE     Rational diversification of a promoter providing fine-tuned 
            expression and orthogonal regulation for synthetic biology.
  JOURNAL   PLoS One. 2012;7(3):e33279. Epub 2012 Mar 19.
  PUBMED    22442681
REFERENCE   2  (bases 1 to 6881)
  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: "PLoS One.";
            date: "2012"; volume: "7(3)"; pages: "e33279. Epub 2012 Mar 19"
COMMENT     Sequence Label: pTAL5-BB
FEATURES             Location/Qualifiers
     source          1..6881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      790..955
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(845..869)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1244..1903)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(1904..2091)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    2242..2745
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2787..2891
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3959..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4448..4467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4701..4718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4819..4913
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        4920..4938
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5022..5463
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5022..5139
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5434..5457
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5502..5522
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5523..5549
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(6315..6722)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     6572..6594
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6586..6603
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6587..6603
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6613..6631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6679..6699)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6679..6697)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6718..6734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6718..6734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6731..6753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6742..6758
                     /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(6766..6772,6773..6790,6791..6796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6811..6832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cctcgtcgcc ttggcctgcc tcggcggacg tcctgccatg gatgcagtga aaaagggatt
       61 gccgcacgcg ccggaattga tcagaagagt caatcgccgt attggcgaac gcacgtccca
      121 tcgcgttgcc gactacgcgc aagtggttcg cgtgctggag tttttccagt gccactccca
      181 cccagcgtac gcatttgatg aggccatgac gcagttcggg atgagcagga acgggttggt
      241 acagctcttt cgcagagtgg gcgtcaccga actcgaagcc cgcggtggaa cgctcccccc
      301 agcctcgcag cgttgggacc gtatcctcca ggcatcaggg atgaaaaggg ccaaaccgtc
      361 ccctacttca gctcaaacac cggatcaggc gtctttgcat gcattcgccg attcgctgga
      421 gcgtgacctt gatgcgccca gcccaatgca cgagggagat cagacgcggg caagcagccg
      481 taaacggtcc cgatcggatc gtgctgtcac cggcccctcc gcacagcagg ctgtcgaggt
      541 gcgcgttccc gaacagcgcg atgcgctgca tttgcccctc agctggaggg taaaacgccc
      601 gcgtaccagg atctggggcg gcctcccgga tccgatatct taataagagc tcttaattaa
      661 caattcttcg ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg
      721 ccagaaattt acgaaaagat ggaaaagggt caaatcgttg gtagatacgt tgttgacact
      781 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
      841 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
      901 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctccaatt
      961 cccggggatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     1021 tatttcacac cgcatagatc cgtcgagttc aagagaaaaa aaaagaaaaa gcaaaaagaa
     1081 aaaaggaaag cgcgcctcgt tcagaatgac acgtatagaa tgatgcatta ccttgtcatc
     1141 ttcagtatca tactgttcgt ataaatactt actgacattc atagctatac atatatacac
     1201 atgtatatat atcgtatgct ggagctttaa ataatcggtg tcactacata agaacacctt
     1261 tggtggaggg aacatcgttg gttccattgg gcgaggtggc ttctcttatg gcaaccgcaa
     1321 gagccttgaa cgcactctca ctacggtgat ggtcattctt gcctcgcaga caatcaacgt
     1381 ggagggtaat tctgcttgcc tctgcaaaac tttcaagaaa atgcgggatc atctcgcaag
     1441 agaggtctcc tactttctcc ctttgcaaac caagttcgac aactgcgtat ggcctgttag
     1501 aaaggtctac caccgctctg gaaagtgcct catccaaagg cgcaaatcct gatccaaacc
     1561 tttttactcc acgcgccagt agagcctctt taaaagcctg accgagagca atcccgcagt
     1621 cttcagtggt gtgatggtcg tctatgtgta agtcaccaat gcactcaacg attagcgacc
     1681 agccggaatg cttggccaga gcatgtatca tgtggtccag aaaccctata cctgtgtgga
     1741 cgttaatcac ttgcgattgt gtggcctgtt ctgctactgc ttctgcctct ttttctggga
     1801 agatcgagtg ctctatcgct aggggaccac cctttaaaga gatagcaatc tgaatcttgg
     1861 tttcatttgt aatacgcttt actagggctt tctgctctgt catctttgcc ttcgtttatc
     1921 ttgcctgctc attttttagt atattcttcg aagaaatcac attactttat ataatgtata
     1981 attcattatg tgataatgcc aatcgctaag aaaaaaaaag agtcatccgc taggtggaaa
     2041 aaaaaaaatg aaaatcatta ccgaggcata aaaaaatata gagtgtacta gaggaggcca
     2101 agagtaatag aaaaagaaaa ttgcgggaaa ggactgtgtt atgacttccc tgactaatgc
     2161 cgtgttcaaa cgatacctgg cagtgactcc tagcgctcac caagctctta aaacgggaat
     2221 ttatggatct taattaagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
     2281 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
     2341 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
     2401 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
     2461 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
     2521 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
     2581 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
     2641 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
     2701 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa gccttagggt
     2761 caggtggcac ttttcgggga aatgtgcgat gaacccctat ttgtttattt ttctaaatac
     2821 attcaaatat gtatccgcta atgagacaat aaccctgata aatgcttcaa ttatattgaa
     2881 aaaggaagag tatgagtatc caacatttcc gtgtcgccct tattcccttt tttgcggcct
     2941 tctgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3001 agttgggtgc ccgagtgggt tacattgaac tggatctcaa cagcggcaaa atcttagaga
     3061 gttttcgccc cgaagaacgc ttcccaatga tgagcacttt caaagttctg ctatgtggcg
     3121 cggtgttatc ccgtattgat gctgggcaag agcaactcgg tcgccgcata cactattctc
     3181 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3241 taagagaatt atgtagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3301 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3361 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3421 acaccacgat gcctgtagca atggcaacaa ctctccgcaa actattaact ggcgaactac
     3481 ttactcttgc ttcacgccag caactcattg actggatgga ggcggacaaa gttgcaggac
     3541 cacttctgcg ctcggcactt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3601 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3661 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3721 agataggtgc ctcactgatt aagcattggt aactgtcaga cgaagggaaa taaatagtag
     3781 cccgcctgat gtgcgggctt ttttttggtg gtggttacca gtgtcgctta gctaccgtag
     3841 gtcctcttag gcggtccgat tagcgacata gtcatcaatg ccttgacggc aaaatccctt
     3901 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac taactctttt tccgaaggta actggcttca
     4081 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gctcacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4621 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4681 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4741 gcaaaccgcc tctccccgcg cgttggccga ttcattgatg caggtcaacg acatcacctg
     4801 gttgcgagcc ttctgaagga ctcctgttga tagatccagt aatgacctca gaactccatc
     4861 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatcaggcgt
     4921 aatacgactc actatagggc gaatggaact ggacgctcgc ctgtggtcgc agggtggctc
     4981 gtacgaactt accgcggaga catatcgatg aattcgaagt acggattaga agccgccgag
     5041 cgggcgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg
     5101 ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata
     5161 ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa
     5221 atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt agccttattt
     5281 ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa
     5341 aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt
     5401 caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag
     5461 gagaaaaaac ccgcctaggt tcaacaatgg cttcctcccc tccaaagaaa aagagaaagg
     5521 ttagttggaa ggacgcaagt ggttggtcta gaatgcatgc ggatcccatt cgtccgcgca
     5581 ggccaagtcc tgcccgcgag cttctgcccg gaccccaacc ggatagggtt cagccgactg
     5641 cagatcgtgg ggtgtctgcg cctgctggca gccctctgga tggcttgccc gctcggcgga
     5701 cggtgtcccg gacccggctg ccatctcccc ctgcgccctc acctgcgttc tcggcgggca
     5761 gcttcagcga tctgctccgt ccgttcgatc cgtcgcttct tgatacatcg cttcttgatt
     5821 cgatgcctgc cgtcggcacg ccgcatacag cggctgcccc agcagagtgg gatgaggcgc
     5881 aatcggctct gcgtgcagcc gatgacccgc cacccaccgt gcgtgtcgct gtcactgccg
     5941 cgcggccgcc gcgcgccaag ccggccccgc gacggcgtgc tgcgcaaccc tccgacgctt
     6001 cgccggccgc gcaggtggat ctacgcacgc tcggctacag tcagcagcag caagagaaga
     6061 tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga ggcactggtg ggccatgggt
     6121 ttacacacgc gcacatcgtt gcgctcagcc aacacccggc agcgttaggg accgtcgctg
     6181 tcacgtatca gcacataatc acggcgttgc cagaggcgac acacgaagac atcgttggcg
     6241 tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt gctcacggat gcgggggagt
     6301 tgagaggtcc gccgttacag ttggacacag gccaacttgt gaagattgca aaacgtggcg
     6361 gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc actgacgggt gcccccctgg
     6421 agacgggcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     6481 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6541 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     6601 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag
     6661 gtcctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     6721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6841 gcgctcactg cccgctttcc accggtcgtc tccaacgacc a
//
