LOCUS       sequence-203955-        9157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses phosphomutant mouse ATP-citrate lyase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9157)
  AUTHORS   Lee JV, Carrer A, Shah S, Snyder NW, Wei S, Venneti S, Worth AJ, 
            Yuan ZF, Lim HW, Liu S, Jackson E, Aiello NM, Haas NB, Rebbeck TR, 
            Judkins A, Won KJ, Chodosh LA, Garcia BA, Stanger BZ, Feldman MD, 
            Blair IA, Wellen KE
  TITLE     Akt-dependent metabolic reprogramming regulates tumor cell histone 
            acetylation.
  JOURNAL   Cell Metab. 2014 Aug 5;20(2):306-19. doi: 
            10.1016/j.cmet.2014.06.004. Epub 2014 Jul 3.
  PUBMED    24998913
REFERENCE   2  (bases 1 to 9157)
  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"; doi: 
            "10.1016/j.cmet.2014.06.004"; journalName: "Cell Metab"; date: 
            "2014-08-5- 5"; volume: "20"; issue: "2"; pages: "306-19"
COMMENT     Sequence Label: pEF6-Acly(S455A)
FEATURES             Location/Qualifiers
     source          1..9157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4575..4604
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4683..4724
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4734..4751
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4774..4791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4780..5004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5050..5478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5137..5156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5347..5368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5487..5507)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5492..5821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5672..5807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5869..5916
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5935..6333
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6491..6612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6528..6547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6582..6601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6661..6677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6661..6677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6674..6696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6685..6701
                     /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(join(6709..6715,6716..6733,6734..6739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6754..6775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6892..6909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7052..7640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7132..7151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7811..8602,8603..8671))
                     /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     8434..8453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8672..8776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8851..8870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt gcccttcccc cgaattccac catgtcagcc aaggcaattt
     1321 cagagcagac cggcaaagaa ctcctgtaca agtacatctg caccacctcg gccatccaga
     1381 accggttcaa gtacgcccgg gtgactcccg acacagactg ggcccatctg ctgcaggacc
     1441 acccgtggct gctcagccag agcttggttg tcaagccaga ccagttaatc aaacgtcgag
     1501 gaaagcttgg cctcgtcggg gtcaatctct ctctggatgg agtcaaatcc tggctaaaac
     1561 ctcgcctggg acatgaagcc actgtcggca aggccaaagg cttcctcaag aactttctca
     1621 ttgaaccctt cgtcccccac agtcaggcgg aggagttcta cgtgtgcatc tatgctaccc
     1681 gggaaggaga ctacgtcctg ttccaccatg aagggggtgt ggatgtgggc gatgtggatg
     1741 ccaaagccca gaagctgctt gtgggtgtgg acgaaaagct gaataccgag gacattaaga
     1801 gacacctgtt ggtccatgca cctgaggaca agaaagaagt cctggccagc ttcatctctg
     1861 gtctattcaa tttctacgag gatctgtact tcacctacct tgagatcaac ccccttgtgg
     1921 tgaccaaaga tggtgtctac atccttgact tggcggccaa ggtggatgcc acagctgact
     1981 acatctgtaa agtcaagtgg ggtgatatag agttccctcc cccctttggg cgtgaggcgt
     2041 accccgagga agcctacatt gcagacctgg atgccaaaag tggagcaagc ttgaagctga
     2101 ccttgctgaa ccccaagggg cggatctgga ccatggttgc tgggggtggc gcctccgtcg
     2161 tgtacagtga caccatctgt gatcttggag gtgtcaatga actggcgaat tacggggaat
     2221 actcgggtgc ccccagtgaa caacagacct atgactatgc caagaccatc ctctcactta
     2281 tgactcgaga gaagcaccca gaaggcaaga tcctcatcat tggaggcagc attgcaaact
     2341 tcaccaatgt ggccgccacc ttcaagggca ttgtgagagc gattcgagat taccagggtc
     2401 ccctgaagga gcatgaggtc accatctttg tccgaagagg tggccccaac tatcaagagg
     2461 gattacgagt gatgggagaa gttgggaaga ccactgggat ccccatccat gtctttggca
     2521 cagaaactca catgacggcc attgtgggca tggccctggg ccaccggccc attcccaacc
     2581 agccacccac agcagctcac actgccaact tcctccttaa tgccagcggg agcacatcga
     2641 ctccagcacc cagtaggaca gcagcttttt ctgagtcccg agctgatgaa gtggcacctg
     2701 caaagaaagc caagccagct atgccccaag gaaagagtgc caccctcttc agccgacata
     2761 ccaaggccat cgtgtggggc atgcagaccc gggctgtgca gggcatgctg gactttgact
     2821 acgtgtgttc ccgagacgag ccctcagtgg ctgccatggt ctaccctttc actggggatc
     2881 acaagcagaa gttttactgg ggacacaagg aaatcctgat ccctgtcttc aagaacatgg
     2941 ctgacgccat gaagaagcac ccggaggtag acgtgctgat caactttgcg tctctgcggt
     3001 ccgcttacga cagcaccatg gagaccatga actatgccca gatccgcacc atagccatca
     3061 tagcagaagg tatccctgag gccctcacac ggaagctcat caagaaggcc gaccagaagg
     3121 gagtgaccat cattgggcca gctacggttg ggggcattaa gcctggatgc tttaagatcg
     3181 ggaatactgg tggaatgctg gacaacatcc tggcctccaa actgtaccgc ccaggcagcg
     3241 tggcctacgt ctcacgttca ggaggcatgt cgaacgagct caataacatc atctctcgga
     3301 ccacagatgg tgtctatgag ggcgtggcca tcggcgggga caggtaccct gggtccacat
     3361 tcatggatca cgtgctgcgc taccaggaca ctccaggagt caagatgatc gtagttcttg
     3421 gggagatagg gggcacagag gaatataaga tctgccgggg catcaaggag ggccgcctca
     3481 ccaagccagt ggtctgctgg tgtatcggga cctgtgccac catgttctcc tccgaggtcc
     3541 agtttggcca tgctggagct tgtgccaacc aggcttctga aactgcagta gccaagaacc
     3601 aggccttgaa ggaagcagga gtgtttgtgc cccgaagctt cgatgagctt ggagaaatca
     3661 ttcagtctgt gtatgaagat ctggtggcca aaggagccat tgtacctgcc caggaagtgc
     3721 cacctccaac agtgcccatg gactactctt gggccagaga gctgggtttg atccgaaaac
     3781 ctgcctcatt catgaccagc atctgtgatg agcgagggca ggagctcatt tatgcgggca
     3841 tgcccatcac cgaggtcttc aaggaggaga tgggcatcgg tggtgtcctc ggcctcctct
     3901 ggttccagag aaggttgccc aagtattcct gccagttcat tgagatgtgt ctgatggtca
     3961 cagctgatca cgggccagct gtctctggag cccataacac catcatctgt gctcgggctg
     4021 ggaaggacct ggtctccagc ctcacctcag ggctgctcac cattggagac cggtttgggg
     4081 gtgccttgga tgccgcagca aagatgttca gtaaagcctt tgacagcggc atcattccca
     4141 tggagttcgt caacaagatg aagaaggagg ggaagctgat catgggcatc ggccatcgag
     4201 taaaatcgat aaacaaccca gacatgcgag tgcagatcct caaggacttc gtcaaacagc
     4261 acttccccgc caccccgctg ctcgactatg ccctggaagt ggagaagatt accacctcca
     4321 agaagccaaa tcttatcctg aatgtggacg gcttcatcgg cgttgcgttt gtggacatgc
     4381 tcaggaactg tggctccttc acccgggagg aagctgatga atatgttgac attggagccc
     4441 tcaatggcat ctttgtgcta ggaaggagta tgggcttcat tgggcactac cttgaccaga
     4501 agaggctgaa gcaagggctg tatcgtcacc cctgggatga catttcctat gttcttccag
     4561 aacacatgag catggaacaa aaactcatct cagaagagga tctgtgaatt cgggggaagg
     4621 gcaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg
     4681 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     4741 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     4801 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4861 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4921 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4981 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5041 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5101 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5161 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5221 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5281 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5341 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5401 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5461 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5521 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5641 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5761 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5821 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     5881 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     5941 aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc
     6001 atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc
     6061 ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg
     6121 ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag
     6181 aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct
     6241 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
     6301 ttgctgccct ctggttatgt gtgggagggc taagcacttc gtggccgagg agcaggactg
     6361 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6421 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6481 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6601 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7021 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7081 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7141 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7201 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7261 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7321 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7381 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7441 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7501 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7561 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7621 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7681 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7741 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7801 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7861 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7921 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7981 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8041 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8101 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8161 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8221 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8281 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8341 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8401 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8461 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8521 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8581 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8641 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8701 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8761 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8821 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     8881 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     8941 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     9001 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     9061 gcttttgcaa aaagctttgc aaagatggat aaagttttaa acagagagga atctttgcag
     9121 ctaatggacc ttctaggtct tgaaaggagt gggaatt
//
