LOCUS       sequence-422472-        6661 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian cell expression vector for overexpression of AnkG ZU5ex 
            isoform in motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6661)
  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 6661)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 ZU5ex-3xHA
FEATURES             Location/Qualifiers
     source          1..6661
                     /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(2296..2315)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2323..3339
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2377..2399)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3340..3616)
                     /label=chicken beta-actin promoter
     enhancer        3618..3997
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4028..4132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4133..4201,4202..4993)
                     /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(4351..4370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5164..5752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5653..5672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5906..5923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5991..6125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6015..6034)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6069..6088
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6131..6326)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6145..6280
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6385..6406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6421..6426,6427..6444,6445..6451)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6459..6475
                     /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     6483..6499
                     /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 tagccttaat
     1621 ccgggaaacc acggcaaaat actgggggaa atcctttgtg ataattctgc agatgcgctt
     1681 tgtacccaac tcttccgggc tgtcgagttc ttcatccatg ccattgagaa gctccgcgag
     1741 gtcttcgttt ttactgtcaa actgatgttc cttccaggtc tctccgttct cgctccgaag
     1801 gacgataagt tctctctcct tccccctcat ggacccaaaa tgagggattt ccacaatgac
     1861 ggggcctaaa aattgtgccc ccgcaggacc catttctacc agcctactgg ctaatccctc
     1921 tccttccacc atggggggtg ggttggccag tttatgtctc tttaccaggc ggcacgtgat
     1981 gcgggtgggg gccgtacact ttcgcggagg gatgatgatc cgcatcccgt ggtggcggct
     2041 tcctcgcatg gagccccctc tcgcgtccac cataaagcta accagaaacc cagaatgcac
     2101 cgggcttgag accaggttca cattatctaa cgtgtccgct gcccaactgt agtgtctgag
     2161 ggagtcagaa tcaaactccc tcgtgaacgt caggtgctgc tctttggatg gtaccagaag
     2221 ttcctctatc atcatgctgt cccttgcgta ggagctagcc atggtaccga attctttgcc
     2281 aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga aaaagaagaa
     2341 ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga agccgaaccc
     2401 cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc gaggctggag
     2461 agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca tttccttcct
     2521 gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc gcctcccaga
     2581 tttcggctcc gcacagattt gggacaaagg aagtccctgc gccctctcgc acgattacca
     2641 taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc gggggccgcc
     2701 gcgcccctcc cccgagccct ccttggcccg aggcggcccc gccccgcccg gcacccccac
     2761 ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgtac ggagccccgc
     2821 acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg gttgcagccc
     2881 gaccgacgcg cccacacccc ctgctcaccc ccccacgcac acaccccgca cgcagccttt
     2941 gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc ccggccgcgc tcccctcgcg
     3001 cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag cgctcacagc cgccgggcag
     3061 cgcggagccg cacgcggcgc tccccacgca cacacacacg cacgcacccc ccgagccgct
     3121 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     3181 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     3241 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     3301 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     3361 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     3421 ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc cccgccccgc
     3481 cccgcctggc gcgcgccccc cccccccccc gcccccatcg ctgcacaaaa taattaaaaa
     3541 ataaataaat acaaaattgg gggtggggag gggggggaga tggggagagt gaagcagaac
     3601 gtggggctca cctcgaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     3661 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     3721 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     3781 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     3841 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     3901 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     3961 attgattact attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg
     4021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5101 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5161 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5281 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5821 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5881 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5941 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg gatccagaca
     6001 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     6061 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     6121 aagttagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     6181 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     6241 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     6301 ctatggttgc tgactaattg agatgcggat ccgctggcac gacaggtttc ccgactggaa
     6361 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6421 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6481 cacaggaaac agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc
     6541 ataagagaag agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta
     6601 gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata
     6661 t
//
