LOCUS       sequence-473491-        6072 bp    DNA     circular SYN 21-MAY-2025
DEFINITION  stable expression of TDH using viral infection .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  AUTHORS   Donato V, Bonora M, Simoneschi D, Sartini D, Kudo Y, Saraf A, 
            Florens L, Washburn MP, Stadtfeld M, Pinton P, Pagano M
  TITLE     The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in 
            mouse embryonic stem cells.
  JOURNAL   Nat Cell Biol. 2017 Apr;19(4):341-351. doi: 10.1038/ncb3491. Epub 
            2017 Mar 20.
  PUBMED    28319092
REFERENCE   2  (bases 1 to 6072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb3491"; 
            journalName: "Nat Cell Biol"; date: "2017-04"; volume: "19"; issue: 
            "4"; pages: "341-351"
COMMENT     Sequence Label: pHAGE2-EF1alpha TDH
FEATURES             Location/Qualifiers
     source          1..6072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      863..1451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1352..1371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1605..1622
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1729..1806
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1807..2889
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1826..1845)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    2902..2923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2938..2943,2944..2961,2962..2968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2976..2992
                     /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     2981..3003
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3000..3016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3000..3016
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3035..3054
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3035..3053
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5135..5153)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5136..5153)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5161..5178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5161..5177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5170..5192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5319..5773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5406..5425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5615..5636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5799..5903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5904..5972,5973..6072,1..692)
                     /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"
ORIGIN
        1 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
       61 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      121 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      181 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      241 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      301 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      361 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      421 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      481 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      541 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      601 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      661 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
      721 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
      781 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
      841 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      901 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      961 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1021 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1081 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1141 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1201 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1261 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1321 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1381 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1441 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1501 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1561 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1621 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1681 aatgcagagc ttgcaattcg cgcgcgaagg cgaagcggca tttacgttga caccatcgaa
     1741 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1801 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1861 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1921 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1981 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     2041 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     2101 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgg
     2161 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     2221 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     2281 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     2341 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     2401 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     2461 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     2521 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     2581 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     2641 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     2701 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     2761 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     2821 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2881 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2941 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     3001 aggaaacagc tatgaccatg attacgccaa gctctaatac gactcactat agggaaagct
     3061 ggtacgcctg caggtaccgg tccggaattc ccgggtcgac aagaggtaat ccttaacatc
     3121 agtgttccca ggaaccagaa acgactaagt tcctagttct tggcggaggt gccaccagac
     3181 acaggtccat agcagatctt tccagctgtg gcttgcaagt cccaaattct aaccccagtc
     3241 aattgtggaa ggggtgtggt ctcgtcttgg tgcagccaca aggtcagctc cgtccgagat
     3301 gtaggtgtgg gcttcccaca agcgtcctgg ggcggaggtg aggtgaggtg aggtaggagg
     3361 gaggcacggc tcccgcggcg gcgaggcggg tccagagcag actgaagata aaaggcagaa
     3421 ccaaactctt gactccgcac gtgctgctcc ggctcccgcg tggcgttctc agcatccagt
     3481 cttctgtgtc tgcagggtat cagaagaaca gatgctattc cttgggatgc taaaacaggt
     3541 ggtcaatggg actgcacaga gcaaagcctc tagctgccgg aagcttgtct tgcccttgaa
     3601 gtttctaggc acctcccagc accggatccc agccgatgcc aacttccact ccacctcgat
     3661 ctcggaagct gagcctcctc gggtcttgat cacagggggt cttggccagc tgggggtagg
     3721 acttgctaac ctcctgagga aacgatttgg gaagaacaat gtgatcctat cagacataag
     3781 gaagccccct gctcatgtct tccacagtgg cccgtttgtt tatgccaaca tcttggatta
     3841 caagagcctc cgtgagatcg tggtgaacca ccgtatcagc tggctgtttc actacagtgc
     3901 cctgctgagt gccgtgggag aggcaaatgt gtccttagcc agagatgtga atataactgg
     3961 attgcataac gttctagatg ttgcagctga atacaacgtg agattgtttg tgcccagcac
     4021 gattggagcc tttggaccta cctctccccg gaaccctgca cccgatctct gtattcaaag
     4081 gcctaggacc atctatgggg tatccaaggt ccacacagag ctcatgggag aatactatta
     4141 ttaccggtac gggttagatt ttcggtgcct gcgttatcct ggaatcattt ctgcagattc
     4201 ccagcctgga ggaggaacaa ctgactacgc cgtccagatt ttccacgctg cagcaaagaa
     4261 cggcacattc gagtgcaacc tcgaggctgg caccaggctc ccaatgatgt acatcagtga
     4321 ctgcctcagg gccaccctgg aggtcatgga agcccctgcc gagcgcctgt ccatgagaac
     4381 ctacaacatc agcgccatga gcttcacccc cgaagagctg gcccaggcgc tccgcaagca
     4441 cgcgcctgac ttccagatca cccactgcgt ggatcccctc cgacaggcca tagcggagag
     4501 ctggcccatg attctagatg acagcaatgc tcggaaggac tggggctgga agcatgactt
     4561 tgatcttcca gagttggtgg ctacgatgct caacttccac ggcgtcagca ccagagttgc
     4621 ccaagtgaac taaagaagag gagcggcccc tgtgtcctca aagtgcaggg cgcacggcag
     4681 tctctaagtt ggagagccag actgaattct agcctctccc aggggacagc tcgcgtgggg
     4741 ttggagttcc tttacacact gcctttgtcc tgtgcctaaa ttggtccaag cacaaaggcc
     4801 agagaaatga agtcctgagt ggtggtggct atctctcggg atctgcagca tttgtattta
     4861 attaaattca atttaagtat cactcctgag acttattctc ttgcccagag accaaagatt
     4921 acatgttagg gttttcaaag ctgttcttca tgtttaagcc tggaatacta attaaaaaaa
     4981 cgatcgcttg caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaagggc ggccgctcta gaggatccaa
     5101 gcttacgtac gcgtgcatgc gacgtcatag ctcttctata gtgtcaccta aattcaattc
     5161 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5221 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5281 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     5341 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5401 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5461 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5521 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5581 tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca aactggaaca
     5641 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5701 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5761 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5821 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5881 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5941 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6001 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6061 agatccttga ga
//
