LOCUS       sequence-356603-        5619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human GFAP (R79C) mutant fused to 3xFLAG tag in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5619)
  AUTHORS   Lee SH, Nam TS, Kim KH, Kim JH, Yoon W, Heo SH, Kim MJ, Shin BA, 
            Perng MD, Choy HE, Jo J, Kim MK, Choi SY
  TITLE     Aggregation-prone GFAP mutation in Alexander disease validated using
            a zebrafish model.
  JOURNAL   BMC Neurol. 2017 Sep 7;17(1):175. doi: 10.1186/s12883-017-0938-7.
  PUBMED    28882119
REFERENCE   2  (bases 1 to 5619)
  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"; journalName: "BMC 
            Neurol."; date: "2017-09-7"; volume: "17(1):175. doi"; pages: " 
            10.1186/s12883-017-0938-7"
COMMENT     Sequence Label: pSYC-190
FEATURES             Location/Qualifiers
     source          1..5619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1287..1310
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1314..1337
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1341..1364
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1419..1475
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(1520..1539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1521..1539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1544..1678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1568..1587)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1622..1641
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1791..1811)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1791..1809)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1830..1846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1830..1846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1843..1865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1854..1870
                     /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(1878..1884,1885..1902,1903..1908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1923..1944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2061..2078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2232..2820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2312..2331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2991..3782,3783..3851))
                     /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     3614..3633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3852..3956)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3982..4437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4119..4140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4331..4350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4508..5492
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        5527..5545
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5527..5544
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tctcctcagg ggagatgatg gtggggggcc tggctcctgg ccgccgtctg ggtcctggca
       61 cccgcctctc cctggctcga atgccccctc cactcccgac ccgggtggat ttctccctgg
      121 ctggggcact caatgctggc ttcaaggaga cccgggccag tgagcgggca gagatgatgg
      181 agctcaatga ctgctttgcc agctacatcg agaaggttcg cttcctggaa cagcaaaaca
      241 aggcgctggc tgctgagctg aaccagctgc gggccaagga gcccaccaag ctggcagacg
      301 tctaccaggc tgagctgcga gagctgcggc tgcggctcga tcaactcacc gccaacagcg
      361 cccggctgga ggttgagagg gacaatctgg cacaggacct ggccactgtg aggcagaagc
      421 tccaggatga aaccaacctg aggctggaag ccgagaacaa cctggctgcc tatagacagg
      481 aagcagatga agccaccctg gcccgtctgg atctggagag gaagattgag tcgctggagg
      541 aggagatccg gttcttgagg aagatccacg aggaggaggt tcgggaactc caggagcagc
      601 tggcccgaca gcaggtccat gtggagcttg acgtggccaa gccagacctc accgcagccc
      661 tgaaagagat ccgcacgcag tatgaggcaa tggcgtccag caacatgcat gaagccgaag
      721 agtggtaccg ctccaagttt gcagacctga cagacgctgc tgcccgcaac gcggagctgc
      781 tccgccaggc caagcacgaa gccaacgact accggcgcca gttgcagtcc ttgacctgcg
      841 acctggagtc tctgcgcggc acgaacgagt ccctggagag gcagatgcgc gagcaggagg
      901 agcggcacgt gcgggaggcg gccagttatc aggaggcgct ggcgcggctg gaggaagagg
      961 ggcagagcct caaggacgag atggcccgcc acttgcagga gtaccaggac ctgctcaatg
     1021 tcaagctggc cctggacatc gagatcgcca cctacaggaa gctgctagag ggcgaggaga
     1081 accggatcac cattcccgtg cagaccttct ccaacctgca gattcgagaa accagcctgg
     1141 acaccaagtc tgtgtcagaa ggccacctca agaggaacat cgtggtgaag accgtggaga
     1201 tgcgggatgg agaggtcatt aaggagtcca agcaggagca caaggatgtg atgatgatat
     1261 ctggcgcgcc taccatgggg cccggcgact acaaggatga cgatgacaag ggcgactaca
     1321 aggatgacga tgacaagggc gactacaagg atgacgatga caagggccgg cctaagatca
     1381 ccggtcccgg gcttaaggag ctcgcatgcg gaagcggagc tactaacttc agcctgctga
     1441 agcaggctgg agacgtggag gagaaccctg gacctagatc tggaattcaa ggcctctcga
     1501 gcctctagat agctagcctc cctatagtga gtcgtattac gtagatccag acatgataag
     1561 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1621 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1681 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1741 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     1801 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1861 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     1921 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1981 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2641 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3961 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4021 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4081 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4141 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4201 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4261 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4321 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4381 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     4441 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4501 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     4561 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     4621 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     4681 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     4741 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     4801 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     4861 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     4921 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     4981 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     5041 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     5101 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     5161 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     5221 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     5281 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     5341 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     5401 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     5461 aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga cgtcggagca
     5521 agcttgattt aggtgacact atagaataca agctacttgt tctttttgca ggatccatgg
     5581 agaggagacg catcacctcc gctgctcgcc gctcctacg
//
