LOCUS       sequence-275427-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged GK5 (isoform 2) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  AUTHORS   Zhang D, Tomisato W, Su L, Sun L, Choi JH, Zhang Z, Wang KW, Zhan X,
            Choi M, Li X, Tang M, Castro-Perez JM, Hildebrand S, Murray AR, 
            Moresco EMY, Beutler B
  TITLE     Skin-specific regulation of SREBP processing and lipid biosynthesis 
            by glycerol kinase 5.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Jun 27;114(26):E5197-E5206. doi: 
            10.1073/pnas.1705312114. Epub 2017 Jun 12.
  PUBMED    28607088
REFERENCE   2  (bases 1 to 6674)
  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: 
            "10.1073/pnas.1705312114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-06-27- 27"; volume: "114"; issue: "26"; pages: 
            "E5197-E5206"
COMMENT     Sequence Label: pcDNA6-C-Flag-GK5_v2
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2562..2585
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2630..2647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2636..2860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2906..3334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2993..3012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3203..3224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3343..3363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3348..3677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3528..3663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3590..3609
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3725..3772
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3791..4189
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4347..4468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4384..4403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4438..4457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4517..4533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4517..4533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4530..4552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4541..4557
                     /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(4565..4571,4572..4589,4590..4595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4610..4631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4748..4765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4919..5507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4999..5018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5678..6469,6470..6538))
                     /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     6301..6320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6539..6643)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 tagagctcct taaggttaac tctagagaat tcggatccat gtcagggcag caggagcgtg
      961 cagagcggca gcgagaggaa ctgagcgcgt cggcctcgcc gccctcccgc ttcgtgctgg
     1021 ggctggacgt gggcagcacg gtgatccgct gccacgtcta tgaccagacg gcgcgagtgc
     1081 gaggctccag cgcgcagaag gtagaaaatg tttaccctca acctggctgg gttgaaatcg
     1141 atcctgattc tctctgggct cagtttgttg ccgtaataaa agatgctgtc aaagctgcag
     1201 gggtgcagat gaatcaaatt gtcggtcttg gcatatcaac acagagagcc actttcatta
     1261 catggaacaa gaaaacaggc catcattttc acaacttcat tagctggcaa gacctaaggg
     1321 cagctgaact tgtaaagtcc tggaataatt ctctcataat gaagctattg catggggcca
     1381 cccgagtcct tcatttcttc agtagaagta aagtaatgct aacggtcagc cgcttcaatt
     1441 tcagcaccca gcatgccacc ttaagattga cctggatttt acaaaaccta tctgaggtga
     1501 aaagggcagt tgaagaagac aattgttgct ttgggacgat tgatacctgg ctgttataca
     1561 agctcacgaa aggttcttcc tacgccacgg attattcaaa tgccagcacc actggatttt
     1621 ttgatcccta tgcgatgcgc tggagcaggc tcataaccac catggtctcc atcccactgt
     1681 ctatcctgcc tcctgtgaag gataccagct ataattttgg atcagtggat gagaagatat
     1741 ttggcgtgcc tataccagtt gttgccttgg ttggtgacca gcagtcggcc atgtttggag
     1801 agtgctgctt cgagacggga gatgtgaagc taaccatggg aactggaaca tttttggata
     1861 tcaatactgg aaaaaacctt cagcatgtta atggaggctt ttatccacta attggatgga
     1921 agattgggca agaacttgta tgtttagctg aaggcaatgc aggagacacg ggcaccgcca
     1981 taatgtgggc tcaaaaatta gatctcttca cagacgctgc tgagactgaa aaaatggccc
     2041 tgagcctgga agattctgaa ggggtttact ttgttccgtc gtttagtgga ttacaggctc
     2101 cattaaatga cccctgtgcc tgtgcctctt ttatgggttt gaagcactcc actaataaat
     2161 accatcttgt ccgagcaata ttggagtcaa tagctttcag gaacaaacag ttgtacgaca
     2221 tgctgcagag agaaatccag attcccgtca cgaacatccg ggcagatgga ggcgtgtgta
     2281 acaacgcttt cgtcatgcag atgacttcag acctgataaa tgagaagata gacagacctg
     2341 cccacttcga catgtcctgc ttgggggcag cctctctggc tggcctcgct gttggatttt
     2401 gggcggacaa ggaagagctc cagaagctga ggcaaagcga gatggttttc aagcctcaga
     2461 agaagtggca ggaatacgag gtgaatatgg aaaactgggt aaaagcggtg aagcgctcca
     2521 tgaattggta taacaagacg ctcgagccat ggaagcttat ggactacaaa gacgatgacg
     2581 acaaggatat ctaactgact gaaccggtgg taccaaaccc gctgatcagc ctcgactgtg
     2641 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2701 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2761 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2821 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2881 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3301 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3601 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3661 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     3721 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3781 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     3841 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     3901 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     3961 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4021 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4081 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4141 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     4201 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4261 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4321 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4381 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4441 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4501 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5641 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5761 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5821 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5881 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6121 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6181 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6361 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6541 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6601 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6661 tgccacctga cgtc
//
