LOCUS       sequence-275421-        6596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GK5 (isoform 1) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6596)
  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 6596)
  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-GK5_v1
FEATURES             Location/Qualifiers
     source          1..6596
                     /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"
     primer_bind     complement(2552..2569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2558..2782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2828..3256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3265..3285)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3270..3599
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3450..3585
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3512..3531
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3647..3694
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3713..4111
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4269..4390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4306..4325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4360..4379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4439..4455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4439..4455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4452..4474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4463..4479
                     /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(4487..4493,4494..4511,4512..4517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4532..4553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4670..4687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4841..5429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4921..4940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5600..6391,6392..6460))
                     /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     6223..6242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6461..6565)
                     /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 gcattttaac tcgagccatg gaagcttgat atctaactga
     2521 ctgaaccggt ggtaccaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2581 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2641 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2701 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2761 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2821 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3181 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3421 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3481 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3541 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3601 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     3661 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     3721 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     3781 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     3841 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     3901 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     3961 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     4021 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     4081 gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag caggactgac
     4141 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4201 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4261 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4321 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4381 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4501 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
