LOCUS       sequence-275424-        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged GK5 (isoform 1) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  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 6620)
  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_v1
FEATURES             Location/Qualifiers
     source          1..6620
                     /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             2508..2531
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2576..2593)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2582..2806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2852..3280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2939..2958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3149..3170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3289..3309)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3294..3623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3474..3609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3536..3555
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3671..3718
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3737..4135
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4293..4414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4330..4349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4384..4403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4463..4479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4463..4479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4476..4498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4487..4503
                     /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(4511..4517,4518..4535,4536..4541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4556..4577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4694..4711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4865..5453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4945..4964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5624..6415,6416..6484))
                     /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     6247..6266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6485..6589)
                     /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 gcatttctcg agccatggaa gcttatggac tacaaagacg
     2521 atgacgacaa ggatatctaa ctgactgaac cggtggtacc aaacccgctg atcagcctcg
     2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2761 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2821 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3061 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3181 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3301 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3541 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3601 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3661 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3721 ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     3781 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     3841 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     3901 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     3961 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     4021 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     4081 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcact
     4141 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     4201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4261 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4321 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4381 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4441 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4501 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4561 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4621 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4681 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4741 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4801 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4861 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4921 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4981 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5041 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5101 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5161 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5221 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5281 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5341 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5401 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5461 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5521 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5581 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5641 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5701 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5761 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5821 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5881 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5941 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6001 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6061 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6121 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6181 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6241 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6301 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6361 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6421 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6481 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6541 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6601 gaaaagtgcc acctgacgtc
//
