LOCUS       sequence-422475-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of AnkB ZU5ex 
            isoform in motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  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 6659)
  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-AnkB ZU5ex-3xHA
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     421..437
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     456..476
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        458..476
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    572..683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     672..689
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    716..1304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             822..833
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1231..1251
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1311..1330)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1313..1445
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(1483..1572)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(1588..1608)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2293..2312)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2320..3336
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2374..2396)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3337..3614)
                     /label=chicken beta-actin promoter
     enhancer        3616..3995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4026..4130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4131..4199,4200..4991)
                     /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(4349..4368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5162..5750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5651..5670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5904..5921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5989..6123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6013..6032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6067..6086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6129..6324)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6143..6278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6383..6404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6419..6424,6425..6442,6443..6449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6457..6473
                     /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)."
     primer_bind     6481..6497
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc
      361 caccaccttc tgataggcag cctgcacctg aggagtgaat tagcggataa caatttcaca
      421 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      481 aaaagctggg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
      541 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
      601 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      661 tggctggcaa ctagaaggca cagtcgaggc tgatcgtacc gagctcgaat tccaggcggg
      721 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
      781 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
      841 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccatggaaag
      901 gacgtcagct tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
      961 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     1021 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     1081 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     1141 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     1201 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     1261 ataccagtca atctttcaca aattttgtaa tccagaggtt gattgtcgac acctgtggag
     1321 agaaaggcaa agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt
     1381 ctctgtctcg acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata
     1441 cttacctgcg gcctcgagtt acttaagacc ggtggcggcc ggagcgtaat ctggaacgtc
     1501 atatggatag gatcctgcat agtccgggac gtcataggga tagcccgcat agtcaggaac
     1561 atcgtatggg taaacggccg caccggtcga ttggaagtag aggttctcgc tagctttgat
     1621 gcgagacacc accgcaaagt actgtgggaa gtcacgagtg atgatgcggc agattcgttt
     1681 cttttccagg tcttctggac tgtccagcac ttcatccatg ccattaagga tttcgttcaa
     1741 ttcatcctca gtgtagtcgc agaaatgctc tttccagctg tccccatttt cactgcgcag
     1801 gaccaccagc tccctctcct ttcctcgaag agcggcgaag tgagggatct ccacgatcac
     1861 gggcccaaga aactgagctc ccgaaggtcc gacttcaatc aggcggctgg ccaggccttc
     1921 tccttccacc atgggcggca ttgttgccag tcgatggcgt ttcacaaggc ggcaggtgac
     1981 gcgagttggg gctgtgcatt tccgaggtgg gataatgatt ctgagtccat tgtgtctgca
     2041 tcctcgcatg gcaccaccac gggcatccac cataaaactg actaggaaac ctgaatgaat
     2101 aggactggaa gaaagagcca cgttgtctaa gttctcagtg ccccagctca gacgataaga
     2161 attcctttct gcctccttgg ctagcgcaga cacctggtgg ctggggatca caaccgtgtc
     2221 gtcaatcatg gcactgtccc tcaggtacga gctagccatg gtaccgaatt ctttgccaaa
     2281 atgatgagac agcacaataa ccagcacgtt gcccaggagc tgtaggaaaa agaagaaggc
     2341 atgaacatgg ttagcagagg ctctagagcc gccggtcaca cgccagaagc cgaaccccgc
     2401 cctgccccgt cccccccgaa ggcagccgtc cccccgcgga cagccccgag gctggagagg
     2461 gagaagggga cggcggcgcg gcgacgcacg aaggccctcc ccgcccattt ccttcctgcc
     2521 ggcgccgcac cgcttcgccc cgcgcccgct agagggggtg cggcggcgcc tcccagattt
     2581 cggctccgca cagatttggg acaaaggaag tccctgcgcc ctctcgcacg attaccataa
     2641 aaggcaatgg ctgcggctcg ccgcgcctcg acagccgccg gcgctccggg ggccgccgcg
     2701 cccctccccc gagccctccc cggcccgagg cggccccgcc ccgcccggca cccccacctg
     2761 ccgccacccc ccgcccggca cggcgagccc cgcgccacgc cccgtacgga gccccgcacc
     2821 cgaagccggg ccgtgctcag caactcgggg aggggggtgc agggggggtt gcagcccgac
     2881 cgacgcgccc acaccccctg ctcacccccc cacgcacaca ccccgcacgc agcctttgtt
     2941 cccctcgcag ccccccccgc accgcggggc accgcccccg gccgcgctcc cctcgcgcac
     3001 actgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc tcacagccgc cgggcagcgc
     3061 ggagccgcac gcggcgctcc ccacgcacac acacacgcac gcaccccccg agccgctccc
     3121 cccgcacaaa gggccctccc ggagcccctc aaggctttca cgcagccaca gaaaagaaac
     3181 aagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt cccgcccgct
     3241 cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg cggcggcgga
     3301 gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc ttcgcttttt
     3361 atagggccgc cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg cgccgctctg
     3421 attggctgcc gccgcacctc tccgcctcgc cccgccccgc ccctcgcccc gccccgcccc
     3481 gcctggcgcg cgcccccccc ccccccccgc ccccatcgct gcacaaaata attaaaaaat
     3541 aaataaatac aaaattgggg gtggggaggg gggggagatg gggagagtga agcagaacgt
     3601 ggggctcacc tcgaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     3661 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     3721 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     3781 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     3841 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3901 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3961 tgattactat taataactag tcaataatca atgtcgaccc aggtggcact tttcggggaa
     4021 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4561 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4621 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4681 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4741 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4801 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4861 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4921 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4981 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5041 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5101 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5821 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5881 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5941 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcgga tccagacatg
     6001 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     6061 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6121 gttagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     6181 tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc
     6241 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact
     6301 atggttgctg actaattgag atgcggatcc gctggcacga caggtttccc gactggaaag
     6361 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6421 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6481 caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgag ggatctccat
     6541 aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa tctggctagt
     6601 aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa acaaaatat
//
