LOCUS       sequence-422470-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of AnkG ZU5in 
            isoform in motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  AUTHORS   Jacko M, Weyn-Vanhentenryck SM, Smerdon JW, Yan R, Feng H, Williams 
            DJ, Pai J, Xu K, Wichterle H, Zhang C
  TITLE     Rbfox Splicing Factors Promote Neuronal Maturation and Axon Initial 
            Segment Assembly.
  JOURNAL   Neuron. 2018 Feb 21;97(4):853-868.e6. doi: 
            10.1016/j.neuron.2018.01.020. Epub 2018 Feb 1.
  PUBMED    29398366
REFERENCE   2  (bases 1 to 6096)
  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: 
            "10.1016/j.neuron.2018.01.020"; journalName: "Neuron"; date: 
            "2018-02-21- 21"; volume: "97"; issue: "4"; pages: "853-868.e6"
COMMENT     Sequence Label: pCAGGS-AnkG ZU5in-3xHA
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     20..36
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     20..36
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     55..75
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        57..75
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    171..282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     271..288
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    315..903
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             421..432
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     830..850
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(910..929)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          912..1044
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(1082..1171)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(1187..1207)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1928..1947)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1955..2971
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2009..2031)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2972..3249)
                     /label=chicken beta-actin promoter
     enhancer        3251..3630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3661..3765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3766..3834,3835..4626)
                     /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     complement(3984..4003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4797..5385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5286..5305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5539..5556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5624..5758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5648..5667)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5702..5721
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(5764..5959)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5778..5913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6018..6039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6054..6059,6060..6077,6078..6084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6092..6096,1..12)
                     /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)."
ORIGIN
        1 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
       61 aaccctcact aaagggaaca aaagctgggc tattgtcttc ccaatcctcc cccttgctgt
      121 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
      181 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      241 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcgtaccg
      301 agctcgaatt ccaggcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag
      361 gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg
      421 attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca
      481 ggcgagcagc catggaaagg acgtcagctt ccccgacaac accacggaat tgtcagtgcc
      541 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
      601 aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca
      661 accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg
      721 ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc
      781 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
      841 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
      901 attgtcgaca cctgtggaga gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta
      961 tcagaaacgc aagagtcttc tctgtctcga caagcccagt ttctattggt ctccttaaac
     1021 ctgtcttgta accttgatac ttacctgcgg cctcgagtta cttaagaccg gtggcggccg
     1081 gagcgtaatc tggaacgtca tatggatagg atcctgcata gtccgggacg tcatagggat
     1141 agcccgcata gtcaggaaca tcgtatgggt aaacggccgc accggtcgat tggaagtaga
     1201 ggttctcgct agccttaatc cgggaaacca cggcaaaata ctgggggaaa tcctttgtga
     1261 taattctgca gatgcgcttt gtacccaact cttccgggct gtcgagttct tcatccatgc
     1321 cattgagaag ctccgcgagg tcttcgtttt tactgtcaaa ctgatgttcc ttccaggtct
     1381 ctccgttctc gctccgaagg acgataagtt ctctctcctt ccccctcatg gacccaaaat
     1441 gagggatttc cacaatgacg gggcctaaaa attgtgcccc cgcaggaccc atttctacca
     1501 gcctactggc taatccctct ccttccacca tggggggtgg gttggccagt ttatgtctct
     1561 ttaccaggcg gcacgtgatg cgggtggggg ccgtacactt tcgcggaggg atgatgatcc
     1621 gcatcccgtg gtggcggctt cctcgcatgg agccccctct cgcgtccacc ataaagctaa
     1681 ccagaaacct tgagtcatac tgaggcagcg gagacgagca gccagaatgc accgggcttg
     1741 agaccaggtt cacattatct aacgtgtccg ctgcccaact gtagtgtctg agggagtcag
     1801 aatcaaactc cctcgtgaac gtcaggtgct gctctttgga tggtaccaga agttcctcta
     1861 tcatcatgct gtcccttgcg taggagctag ccatggtacc gaattctttg ccaaaatgat
     1921 gagacagcac aataaccagc acgttgccca ggagctgtag gaaaaagaag aaggcatgaa
     1981 catggttagc agaggctcta gagccgccgg tcacacgcca gaagccgaac cccgccctgc
     2041 cccgtccccc ccgaaggcag ccgtcccccc gcggacagcc ccgaggctgg agagggagaa
     2101 ggggacggcg gcgcggcgac gcacgaaggc cctccccgcc catttccttc ctgccggcgc
     2161 cgcaccgctt cgccccgcgc ccgctagagg gggtgcggcg gcgcctccca gatttcggct
     2221 ccgcacagat ttgggacaaa ggaagtccct gcgccctctc gcacgattac cataaaaggc
     2281 aatggctgcg gctcgccgcg cctcgacagc cgccggcgct ccgggggccg ccgcgcccct
     2341 cccccgagcc ctccccggcc cgaggcggcc ccgccccgcc cggcaccccc acctgccgcc
     2401 accccccgcc cggcacggcg agccccgcgc cacgccccgt acggagcccc gcacccgaag
     2461 ccgggccgtg ctcagcaact cggggagggg ggtgcagggg gggttgcagc ccgaccgacg
     2521 cgcccacacc ccctgctcac ccccccacgc acacaccccg cacgcagcct ttgttcccct
     2581 cgcagccccc cccgcaccgc ggggcaccgc ccccggccgc gctcccctcg cgcacactgc
     2641 ggagcgcaca aagccccgcg ccgcgcccgc agcgctcaca gccgccgggc agcgcggagc
     2701 cgcacgcggc gctccccacg cacacacaca cgcacgcacc ccccgagccg ctccccccgc
     2761 acaaagggcc ctcccggagc ccctcaaggc tttcacgcag ccacagaaaa gaaacaagcc
     2821 gtcattaaac caagcgctaa ttacagcccg gaggagaagg gccgtcccgc ccgctcacct
     2881 gtgggagtaa cgcggtcagt cagagccggg gcgggcggcg cgaggcggcg gcggagcggg
     2941 gcacggggcg aaggcagcgc gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg
     3001 gccgccgccg ccgccgcctc gccataaaag gaaactttcg gagcgcgccg ctctgattgg
     3061 ctgccgccgc acctctccgc ctcgccccgc cccgcccctc gccccgcccc gccccgcctg
     3121 gcgcgcgccc cccccccccc cccgccccca tcgctgcaca aaataattaa aaaataaata
     3181 aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag aacgtggggc
     3241 tcacctcgac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3301 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3361 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3421 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     3481 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     3541 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     3601 actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg
     3661 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3721 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3781 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3841 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3901 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3961 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4021 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4081 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4141 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4201 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4261 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4321 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4381 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4441 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4501 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4561 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4621 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4681 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4741 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4801 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4861 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4921 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4981 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5041 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5101 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5161 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5221 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5281 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5341 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5401 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5461 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5521 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5581 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcggatccag acatgataag
     5641 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5701 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttag
     5761 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5821 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5881 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5941 tgctgactaa ttgagatgcg gatccgctgg cacgacaggt ttcccgactg gaaagcgggc
     6001 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6061 tttatgcttc cggctcgtat gttgtgtgga attgtg
//
