LOCUS       Exported                7423 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of PGC-1a with myc and his tags.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA4 myc PGC-1 alpha
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  AUTHORS   Ichida M, Nemoto S, Finkel T
  TITLE     Identification of a specific molecular repressor of the peroxisome 
            proliferator-activated receptor gamma Coactivator-1 alpha 
            (PGC-1alpha).
  JOURNAL   J Biol Chem. 2002 Dec 27. 277(52):50991-5.
  PUBMED    12397057
REFERENCE   2  (bases 1 to 7423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3348..3377
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3393..3410
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3433..3450)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3439..3663
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3709..4137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3796..3815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4006..4027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4146..4166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4151..4480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4331..4466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4393..4412
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4528..4575
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4594..4968
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5098..5219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5135..5154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5189..5208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5268..5284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5281..5303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5292..5308
                     /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(5316..5346)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5499..5516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6429..7289)
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gttaagcttg gtaccgagct cggatccgga gctggatggc gtgggacatg tgcaaccagg
      961 actctgagtc tgtatggagt gacatcgagt gtgctgctct ggttggtgaa gaccagcctc
     1021 tttgcccaga tcttcctgaa cttgatcttt ctgaactaga tgtgaacgac ttggatacag
     1081 acagctttct gggtggactc aagtggtgca gtgaccaatc agaaataata tccaatcagt
     1141 acaacaatga gccttcaaac atatttgaga agatagatga agagaatgag gcaaacttgc
     1201 tagcagtcct cacagagaca ctagacagtc tccctgtgga tgaagacgga ttgccctcat
     1261 ttgatgcgct gacagatgga gacgtgacca ctgacaatga ggctagtcct tcctccatgc
     1321 ctgacggcac ccctccaccc caggaggcag aagagccgtc tctacttaag aagctcttac
     1381 tggcaccagc caacactcag ctaagttata atgaatgcag tggtctcagt acccagaacc
     1441 atgcaaatca caatcacagg atcagaacaa accctgcaat tgttaagact gagaattcat
     1501 ggagcaataa agcgaagagt atttgtcaac agcaaaagcc acaaagacgt ccctgctcgg
     1561 agcttctcaa atatctgacc acaaacgatg accctcctca caccaaaccc acagagaaca
     1621 gaaacagcag cagagacaaa tgcacctcca aaaagaagtc ccacacacag tcgcagtcac
     1681 aacacttaca agccaaacca acaactttat ctcttcctct gaccccagag tcaccaaatg
     1741 accccaaggg ttccccattt gagaacaaga ctattgaacg caccttaagt gtggaactct
     1801 ctggaactgc aggcctaact ccacccacca ctcctcctca taaagccaac caagataacc
     1861 cttttagggc ttctccaaag ctgaagtcct cttgcaagac tgtggtgcca ccaccatcaa
     1921 agaagcccag gtacagtgag tcttctggta cacaaggcaa taactccacc aagaaagggc
     1981 cggagcaatc cgagttgtat gcacaactca gcaagtcctc agtcctcact ggtggacacg
     2041 aggaaaggaa gaccaagcgg cccagtctgc ggctgtttgg tgaccatgac tattgccagt
     2101 caattaattc caaaacggaa atactcatta atatatcaca ggagctccaa gactctagac
     2161 aactagaaaa taaagatgtc tcctctgatt ggcaggggca gatttgttct tccacagatt
     2221 cagaccagtg ctacctgaga gagactttgg aggcaagcaa gcaggtctct ccttgcagca
     2281 caagaaaaca gctccaagac caggaaatcc gagccgagct gaacaagcac ttcggtcatc
     2341 ccagtcaagc tgtttttgac gacgaagcag acaagaccag tgaactgagg gacagtgatt
     2401 tcagtaatga acaattctcc aaactaccta tgtttataaa ttcaggacta gccatggatg
     2461 gcctgtttga tgacagcgaa gatgaaagtg ataaactgag ctacccttgg gatggcacac
     2521 aatcctattc attgttcaat gtgtctcctt cttgttcttc ttttaactct ccatgtagag
     2581 attctgtgtc accacccaaa tccttatttt ctcaaagacc ccaaaggatg cgctctcgtt
     2641 caaggtcctt ttctcgacac aggtcgtgtt cccgatcacc atattccagg tcaagatcaa
     2701 ggtctccagg cagtagatcc tcttcaagat cctgctatta ctatgagtca agccactaca
     2761 gacaccgcac gcaccgaaat tctcccttgt atgtgagatc acgttcaaga tcgccctaca
     2821 gccgtcggcc caggtatgac agctacgagg aatatcagca cgagaggctg aagagggaag
     2881 aatatcgcag agagtatgag aagcgagagt ctgagagggc caagcaaagg gagaggcaga
     2941 ggcagaaggc aattgaagag cgccgtgtga tttatgtcgg taaaatcaga cctgacacaa
     3001 cacggacaga actgagggac cgttttgaag tttttggtga aattgaggag tgcacagtaa
     3061 atctgcggga tgatggagac agctatggtt tcattaccta ccgttatacc tgtgatgctt
     3121 ttgctgctct tgaaaatgga tacactttgc gcaggtcaaa cgaaactgac tttgagctgt
     3181 acttttgtgg acgcaagcaa tttttcaagt ctaactatgc agacctagat tcaaactcag
     3241 atgactttga ccctgcttcc accaagagca agtatgactc tctggatttt gatagtttac
     3301 tgaaagaagc tcagagaagc ttgcgcaggt ctcgaggtca cccattcgaa caaaaactca
     3361 tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat tgagtttaaa
     3421 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3481 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3541 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3601 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3661 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3721 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3781 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3841 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3901 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3961 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4021 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4081 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4141 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4201 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4321 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4381 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4441 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4501 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     4561 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     4621 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     4681 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     4741 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     4801 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     4861 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     4921 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     4981 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5041 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5221 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acg
//
