LOCUS       Exported                6828 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTAL6-BB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  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 6828)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6828
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      794..959
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(849..873)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1248..1907)
                     /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(1908..2095)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    2246..2749
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2804..2895
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3963..4551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4452..4471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4705..4722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4823..4917
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        4924..4942
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5019..5418
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5453..5473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5474..5500
                     /codon_start=1
                     /product="epitope tag from the baculovirus Autographa 
                     californica GP64 envelope fusion protein"
                     /label=AcV5 tag
                     /translation="SWKDASGWS"
     CDS             complement(6266..6673)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     6523..6545
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6537..6554
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6538..6554
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6564..6582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6630..6650)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6630..6648)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6669..6685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6669..6685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6682..6704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6693..6709
                     /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(6717..6747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6762..6783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 accacctcgt cgccttggcc tgcctcggcg gacgtcctgc catggatgca gtgaaaaagg
       61 gattgccgca cgcgccggaa ttgatcagaa gagtcaatcg ccgtattggc gaacgcacgt
      121 cccatcgcgt tgccgactac gcgcaagtgg ttcgcgtgct ggagtttttc cagtgccact
      181 cccacccagc gtacgcattt gatgaggcca tgacgcagtt cgggatgagc aggaacgggt
      241 tggtacagct ctttcgcaga gtgggcgtca ccgaactcga agcccgcggt ggaacgctcc
      301 ccccagcctc gcagcgttgg gaccgtatcc tccaggcatc agggatgaaa agggccaaac
      361 cgtcccctac ttcagctcaa acaccggatc aggcgtcttt gcatgcattc gccgattcgc
      421 tggagcgtga ccttgatgcg cccagcccaa tgcacgaggg agatcagacg cgggcaagca
      481 gccgtaaacg gtcccgatcg gatcgtgctg tcaccggccc ctccgcacag caggctgtcg
      541 aggtgcgcgt tcccgaacag cgcgatgcgc tgcatttgcc cctcagctgg agggtaaaac
      601 gcccgcgtac caggatctgg ggcggcctcc cggatccgat atcttaataa gagctcttaa
      661 ttaacaattc ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac
      721 cttgccagaa atttacgaaa agatggaaaa gggtcaaatc gttggtagat acgttgttga
      781 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
      841 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
      901 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctcc
      961 aattcccggg gattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     1021 gcggtatttc acaccgcata gatccgtcga gttcaagaga aaaaaaaaga aaaagcaaaa
     1081 agaaaaaagg aaagcgcgcc tcgttcagaa tgacacgtat agaatgatgc attaccttgt
     1141 catcttcagt atcatactgt tcgtataaat acttactgac attcatagct atacatatat
     1201 acacatgtat atatatcgta tgctggagct ttaaataatc ggtgtcacta cataagaaca
     1261 cctttggtgg agggaacatc gttggttcca ttgggcgagg tggcttctct tatggcaacc
     1321 gcaagagcct tgaacgcact ctcactacgg tgatggtcat tcttgcctcg cagacaatca
     1381 acgtggaggg taattctgct tgcctctgca aaactttcaa gaaaatgcgg gatcatctcg
     1441 caagagaggt ctcctacttt ctccctttgc aaaccaagtt cgacaactgc gtatggcctg
     1501 ttagaaaggt ctaccaccgc tctggaaagt gcctcatcca aaggcgcaaa tcctgatcca
     1561 aaccttttta ctccacgcgc cagtagagcc tctttaaaag cctgaccgag agcaatcccg
     1621 cagtcttcag tggtgtgatg gtcgtctatg tgtaagtcac caatgcactc aacgattagc
     1681 gaccagccgg aatgcttggc cagagcatgt atcatgtggt ccagaaaccc tatacctgtg
     1741 tggacgttaa tcacttgcga ttgtgtggcc tgttctgcta ctgcttctgc ctctttttct
     1801 gggaagatcg agtgctctat cgctagggga ccacccttta aagagatagc aatctgaatc
     1861 ttggtttcat ttgtaatacg ctttactagg gctttctgct ctgtcatctt tgccttcgtt
     1921 tatcttgcct gctcattttt tagtatattc ttcgaagaaa tcacattact ttatataatg
     1981 tataattcat tatgtgataa tgccaatcgc taagaaaaaa aaagagtcat ccgctaggtg
     2041 gaaaaaaaaa aatgaaaatc attaccgagg cataaaaaaa tatagagtgt actagaggag
     2101 gccaagagta atagaaaaag aaaattgcgg gaaaggactg tgttatgact tccctgacta
     2161 atgccgtgtt caaacgatac ctggcagtga ctcctagcgc tcaccaagct cttaaaacgg
     2221 gaatttatgg atcttaatta agacggatcg cttgcctgta acttacacgc gcctcgtatc
     2281 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
     2341 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
     2401 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
     2461 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
     2521 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
     2581 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
     2641 tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat
     2701 atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg aaaagcctta
     2761 gggtcaggtg gcacttttcg gggaaatgtg cgatgaaccc ctatttgttt atttttctaa
     2821 atacattcaa atatgtatcc gctaatgaga caataaccct gataaatgct tcaattatat
     2881 tgaaaaagga agagtatgag tatccaacat ttccgtgtcg cccttattcc cttttttgcg
     2941 gccttctgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3001 gatcagttgg gtgcccgagt gggttacatt gaactggatc tcaacagcgg caaaatctta
     3061 gagagttttc gccccgaaga acgcttccca atgatgagca ctttcaaagt tctgctatgt
     3121 ggcgcggtgt tatcccgtat tgatgctggg caagagcaac tcggtcgccg catacactat
     3181 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     3241 acagtaagag aattatgtag tgctgccata accatgagtg ataacactgc ggccaactta
     3301 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3421 cgtgacacca cgatgcctgt agcaatggca acaactctcc gcaaactatt aactggcgaa
     3481 ctacttactc ttgcttcacg ccagcaactc attgactgga tggaggcgga caaagttgca
     3541 ggaccacttc tgcgctcggc acttccggct ggctggttta ttgctgataa atctggagcc
     3601 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3661 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3721 gctgagatag gtgcctcact gattaagcat tggtaactgt cagacgaagg gaaataaata
     3781 gtagcccgcc tgatgtgcgg gctttttttt ggtggtggtt accagtgtcg cttagctacc
     3841 gtaggtcctc ttaggcggtc cgattagcga catagtcatc aatgccttga cggcaaaatc
     3901 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3961 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4021 ccagcggtgg tttgtttgcc ggatcaagag ctactaactc tttttccgaa ggtaactggc
     4081 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4261 aaggcgcagc ggtcgggctg aacggggggt tcgtgctcac agcccagctt ggagcgaacg
     4321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4621 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4681 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4741 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt gatgcaggtc aacgacatca
     4801 cctggttgcg agccttctga aggactcctg ttgatagatc cagtaatgac ctcagaactc
     4861 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcag
     4921 gcgtaatacg actcactata gggcgaatgg aactggacgc tcgcctgtgg tcgcagggtg
     4981 gctcgtacga acttaccgcg gagacatatc gatgaattca tagcttcaaa atgtttctac
     5041 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     5101 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     5161 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     5221 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     5281 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     5341 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     5401 atagcaatct aatctaagtt tccgcctagg ttcaacaatg gcttcctccc ctccaaagaa
     5461 aaagagaaag gttagttgga aggacgcaag tggttggtct agaatgcatg cggatcccat
     5521 tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt
     5581 tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc
     5641 cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt
     5701 ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc
     5761 gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg
     5821 ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc
     5881 tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc
     5941 ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca
     6001 gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt
     6061 gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg
     6121 gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga
     6181 catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga
     6241 tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc
     6301 aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg
     6361 tgcccccctg gagacgggcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     6421 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     6481 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     6541 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
     6601 cccccctcga ggtcctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     6661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6781 ttaattgcgt tgcgctcact gcccgctttc caccggtcgt ctccaacg
//
