LOCUS       sequence-41260-e        7242 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 7242)
  AUTHORS   Gruning NM, Rinnerthaler M, Bluemlein K, Mulleder M, Wamelink MM, 
            Lehrach H, Jakobs C, Breitenbach M, Ralser M
  TITLE     Pyruvate kinase triggers a metabolic feedback loop that controls 
            redox metabolism in respiring cells.
  JOURNAL   Cell Metab. 2011 Sep 7;14(3):415-27.
  PUBMED    21907146
REFERENCE   2  (bases 1 to 7242)
  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: "Cell 
            Metab."; date: "2011-09-7"; volume: "14(3)"; pages: "415-27"
COMMENT     Sequence Label: p416GPD-PYK1
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2800..3443
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3337..3360
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3449..3465
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3449..3465
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4982..5229
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4998..5016)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5247..5266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5248..5266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5276..5293)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5276..5292)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5285..5307)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5437..5892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5524..5543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5734..5755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6023..6826)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6827..7047)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7073..7092)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7192..7214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcagtttat cattatcaat actcgccatt
     2821 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     2881 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     2941 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     3001 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     3061 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     3121 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     3181 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     3241 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     3301 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     3361 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     3421 aaaacaccag aacttagttt cgacggattc tagaactagt ggatccatgt ctagattaga
     3481 aagattgacc tcattaaacg ttgttgctgg ttctgacttg agaagaacct ccatcattgg
     3541 taccatcggt ccaaagacca acaacccaga aaccttggtt gctttgagaa aggctggttt
     3601 gaacattgtc cgtatgaact tctctcacgg ttcttacgaa taccacaagt ctgtcattga
     3661 caacgccaga aagtccgaag aattgtaccc aggtagacca ttggccattg ctttggacac
     3721 caagggtcca gaaatcagaa ctggtaccac caccaacgat gttgactacc caatcccacc
     3781 aaaccacgaa atgatcttca ccaccgatga caagtacgct aaggcttgtg acgacaagat
     3841 catgtacgtt gactacaaga acatcaccaa ggtcatctcc gctggtagaa tcatctacgt
     3901 tgatgatggt gttttgtctt tccaagtttt ggaagtcgtt gacgacaaga ctttgaaggt
     3961 caaggctttg aacgccggta agatctgttc ccacaagggt gtcaacttac caggtaccga
     4021 tgtcgatttg ccagctttgt ctgaaaagga caaggaagat ttgagattcg gtgtcaagaa
     4081 cggtgtccac atggtcttcg cttctttcat cagaaccgcc aacgatgttt tgaccatcag
     4141 agaagtcttg ggtgaacaag gtaaggacgt caagatcatt gtcaagattg aaaaccaaca
     4201 aggtgttaac aacttcgacg aaatcttgaa ggtcactgac ggtgttatgg ttgccagagg
     4261 tgacttgggt attgaaatcc cagccccaga agtcttggct gtccaaaaga aattgattgc
     4321 taagtctaac ttggctggta agccagttat ctgtgctacc caaatgttgg aatccatgac
     4381 ttacaaccca agaccaacca gagctgaagt ttccgatgtc ggtaacgcta tcttggatgg
     4441 tgctgactgt gttatgttgt ctggtgaaac cgccaagggt aactacccaa tcaacgccgt
     4501 taccactatg gctgaaaccg ctgtcattgc tgaacaagct atcgcttact tgccaaacta
     4561 cgatgacatg agaaactgta ctccaaagcc aacctccacc accgaaaccg tcgctgcctc
     4621 cgctgtcgct gctgttttcg aacaaaaggc caaggctatc attgtcttgt ccacttccgg
     4681 taccacccca agattggttt ccaagtacag accaaactgt ccaatcatct tggttaccag
     4741 atgcccaaga gctgctagat tctctcactt gtacagaggt gtcttcccat tcgttttcga
     4801 aaaggaacct gtctctgact ggactgatga tgttgaagcc cgtatcaact tcggtattga
     4861 aaaggctaag gaattcggta tcttgaagaa gggtgacact tacgtttcca tccaaggttt
     4921 caaggccggt gctggtcact ccaacacttt gcaagtctct accgtttaag tcgacctcga
     4981 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     5041 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     5101 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     5161 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     5221 ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     5281 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5341 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5401 cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa
     5461 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5521 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5581 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5641 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5701 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5761 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5821 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5881 cgtttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     5941 catagggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     6001 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     6061 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     6121 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     6181 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     6241 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     6301 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     6361 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     6421 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     6481 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     6541 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     6601 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     6661 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     6721 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     6781 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     6841 tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     6901 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     6961 tctgttcgga gattaccgaa tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat
     7021 aaaaaaaaaa tgatgaattg aattgaaaag ctgtggtatg gtgcactctc agtacaatct
     7081 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     7141 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     7201 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
