LOCUS       sequence-43492-e        7078 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 7078)
  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 7078)
  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: p413TEF-PYK2
FEATURES             Location/Qualifiers
     source          1..7078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..28)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10..28)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(38..55)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(38..54)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(47..69)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      199..654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(286..305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     496..517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(914..1573)
                     /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(1574..1760)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(1907..1926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2026..2048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2086..2104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2146..2649
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2686..2790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2791..2859,2860..3651)
                     /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(3009..3028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3822..4410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4311..4330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4564..4581
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4734..4739,4740..4757,4758..4764)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4772..4788
                     /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     4777..4799
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4796..4812
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4796..4812
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4831..4851
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4833..4851
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4869..5267
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     5271..5287
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5271..5287
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      6822..7069
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6838..6856)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
       61 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      121 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      181 tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      241 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
      301 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
      361 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
      421 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
      481 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
      541 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
      601 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat
      661 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagatc cgtcgagttc
      721 aagagaaaaa aaaagaaaaa gcaaaaagaa aaaaggaaag cgcgcctcgt tcagaatgac
      781 acgtatagaa tgatgcatta ccttgtcatc ttcagtatca tactgttcgt atacatactt
      841 actgacattc ataggtatac atatatacac atgtatatat atcgtatgct gcagctttaa
      901 ataatcggtg tcactacata agaacacctt tggtggaggg aacatcgttg gtaccattgg
      961 gcgaggtggc ttctcttatg gcaaccgcaa gagccttgaa cgcactctca ctacggtgat
     1021 gatcattctt gcctcgcaga caatcaacgt ggagggtaat tctgctagcc tctgcaaagc
     1081 tttcaagaaa atgcgggatc atctcgcaag agagatctcc tactttctcc ctttgcaaac
     1141 caagttcgac aactgcgtac ggcctgttcg aaagatctac caccgctctg gaaagtgcct
     1201 catccaaagg cgcaaatcct gatccaaacc tttttactcc acgcgccagt agggcctctt
     1261 taaaagcttg accgagagca atcccgcagt cttcagtggt gtgatggtcg tctatgtgta
     1321 agtcaccaat gcactcaacg attagcgacc agccggaatg cttggccaga gcatgtatca
     1381 tatggtccag aaaccctata cctgtgtgga cgttaatcac ttgcgattgt gtggcctgtt
     1441 ctgctactgc ttctgcctct ttttctggga agatcgagtg ctctatcgct aggggaccac
     1501 cctttaaaga gatcgcaatc tgaatcttgg tttcatttgt aatacgcttt actagggctt
     1561 tctgctctgt catctttgcc ttcgtttatc ttgcctgctc attttttagt atattcttcg
     1621 aagaaatcac attactttat ataatgtata attcattatg tgataatgcc aatcgctaag
     1681 aaaaaaaaag agtcatccgc taggggaaaa aaaaaaatga aaatcattac cgaggcataa
     1741 aaaaatatag agtgtactag aggaggccaa gagtaataga aaaagaaaat tgcgggaaag
     1801 gactgtgtta tgacttccct gactaatgcc gtgttcaaac gatacctggc agtgactcct
     1861 agcgctcacc aagctcttaa aacgggaatt tatggtgcac tctcagtaca atctgctctg
     1921 atgccgcata gttaagccag ccccgacacc cgccaacacg cgctgacgcg ccctgacggg
     1981 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2041 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     2101 tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta
     2161 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
     2221 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
     2281 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
     2341 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
     2401 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
     2461 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
     2521 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
     2581 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
     2641 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2701 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2761 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2821 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2881 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2941 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3001 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3061 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3121 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3181 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3241 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3301 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3361 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3421 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3481 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3541 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3601 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3661 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3721 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3781 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3901 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3961 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4021 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4081 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4141 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4201 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4261 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4321 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4381 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4441 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4501 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4561 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4621 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4681 agtgagcgca acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac
     4741 tttatgcttc cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4801 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     4861 ctggagctca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
     4921 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
     4981 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
     5041 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
     5101 cttgaaaatt ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt
     5161 aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt
     5221 ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta
     5281 gtggatccat gccagagtcc agattgcaga gactagctaa tttgaaaata ggaactccgc
     5341 agcagctcag acgcacctcc ataataggta ccattgggcc caagacaaat agctgcgagg
     5401 ccattactgc tctgagaaaa gctggtttga acatcattcg attgaacttt tcccatggct
     5461 cctacgaatt ccatcaatca gtaatcgaaa atgctgtgaa atcggaacag caattccctg
     5521 gcaggccgct cgccattgcc ctggatacca agggtcccga gatcagaaca ggtcgcacgt
     5581 taaatgacca agatctttat atccccgtag accaccaaat gatctttacc actgacgcaa
     5641 gttttgcaaa cacctccaat gataaaatca tgtatataga ctatgctaac ctgacaaaag
     5701 ttatcgttcc ggggagattt atatacgtgg acgacgggat tctctctttt aaagtgctcc
     5761 aaatcattga cgaatctaat ttaagggtgc aagcggtaaa ctcgggttat atcgcatctc
     5821 ataaaggtgt taatctgcct aataccgacg ttgatttgcc ccccttgtcc gccaaagaca
     5881 tgaaggactt gcaattcgga gtccgcaatg gcattcacat cgtatttgcc tctttcataa
     5941 gaacttcaga agatgtgttg tctatcagaa aagcgttggg ttctgaaggg caagatatca
     6001 agattatatc caagatagaa aaccagcaag ggttggataa ttttgacgaa atcctggaag
     6061 tcacggatgg tgttatgata gcgagaggcg atttaggaat tgaaatcctg gcacctgaag
     6121 tattagccat tcaaaaaaag ctgattgcaa aatgtaattt ggcgggcaaa cctgtcattt
     6181 gcgcgactca gatgctggat tcaatgacac acaatccgag accgacaagg gctgaagtat
     6241 cggatgtggg taacgctgtg ttggatggtg ctgattgtgt tatgctttct ggagaaacgg
     6301 cgaagggtga ttatccggtg aatgcagtta atattatggc ggcgaccgct ctgattgctg
     6361 aaagtactat cgctcatttg gctctttatg acgatctcag agacgccact cccaaaccta
     6421 cttccactac ggaaactgta gcagctgcag ctaccgcagc aatcttggag caagatggta
     6481 aggccatcgt tgtattatct actacaggga acacggcaag gctactgtcg aagtatagac
     6541 caagctgccc tatcatatta gtaacaagac acgcaagaac ggcaagaatt gcgcatttgt
     6601 atagaggtgt tttcccattt ctgtatgaac cgaaacgcct agacgactgg ggtgaggatg
     6661 ttcataggcg cctaaagttt ggtgttgaaa tggcgaggtc tttcggaatg gtggacaacg
     6721 gtgatactgt tgtttccatt caaggattca aaggaggagt cggccattcc aataccttac
     6781 gcatttctac tgttggtcaa gaattctagg tcgacctcga gtcatgtaat tagttatgtc
     6841 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     6901 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     6961 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     7021 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtac
//
