LOCUS       sequence-275422-        6647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GK5 (isoform 2) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  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 6647)
  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_v2
FEATURES             Location/Qualifiers
     source          1..6647
                     /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(2603..2620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2609..2833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2879..3307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2966..2985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3176..3197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3316..3336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3321..3650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3501..3636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3563..3582
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3698..3745
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3764..4162
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4320..4441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4357..4376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4411..4430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4490..4506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4490..4506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4503..4525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4514..4530
                     /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(4538..4544,4545..4562,4563..4568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4583..4604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4721..4738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4892..5480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4972..4991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5651..6442,6443..6511))
                     /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     6274..6293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6512..6616)
                     /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 ggaagcttga tatctaactg actgaaccgg
     2581 tggtaccaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2641 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2701 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2761 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2821 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2881 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2941 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3001 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3061 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3181 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3241 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3301 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3661 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     3721 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc
     3781 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     3841 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     3901 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     3961 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     4021 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     4081 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     4141 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac
     4201 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4261 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4321 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4381 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4441 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4501 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4561 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4621 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4681 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4741 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4801 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4861 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4921 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4981 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5041 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5101 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5161 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5221 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5281 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5341 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5401 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5461 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5521 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5581 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5641 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5701 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5761 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5821 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5881 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5941 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6001 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6061 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6121 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6181 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6241 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6301 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6361 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6421 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6481 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6541 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6601 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
