LOCUS       sequence-422465-        7122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of bIV spectrin 
            (repeats 13-15) in NIH/3T3 cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7122)
  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 7122)
  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-3xFLAG-bIV spectrin (repeats 13-15)
FEATURES             Location/Qualifiers
     source          1..7122
                     /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"
     regulatory      2313..2322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2614..2634)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2641..2712)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2755..2774)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2782..3798
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2836..2858)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3799..4077)
                     /label=chicken beta-actin promoter
     enhancer        4079..4458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4594..4662,4663..5454)
                     /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(4812..4831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5625..6213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6114..6133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6367..6384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6452..6586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6476..6495)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6530..6549
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6592..6787)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6606..6741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6846..6867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6882..6887,6888..6905,6906..6912)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6920..6936
                     /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     6944..6960
                     /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 acttaaggat aagctgctcc acctcatcca cactgctgcc
     1501 cagctcccgg ctctgcagga gcggttcctg agctgtcagc caggcgtcag ccaccacggc
     1561 ctcctgagca aactggtgta cctccagcat ctgctgcagc cactcccagt gacggtccca
     1621 cttctccgac acctcctcct tccggcttcc cagcttgtcc agctgggcct ggatctcatc
     1681 agccatggcg cttttgttga gtaatagaga ccgccccagc tcctggcagg ttgccagctc
     1741 aggcacgcgc gcctccaact ctgtcttcag gccctggtgg tagttcataa gcacctccac
     1801 cgatgacaca tccctgggtt tgtcagctgc cccgatctgg cccgcgatgc cgtccatcca
     1861 ggagagcagg tcgcgggcct ggctgtggaa gcgcagggcg tcggccgtgg agctgacgtg
     1921 cagacgagca tcttcacagg ctgctagcag ctccttccag ccctgcaaca cctcctgctc
     1981 gcggctggcg atggcctcgg cgtgctcgcc agcgtacacc gtccgcagct gggccgcccc
     2041 ttcctgcagc tgccgtacct gggacacgag gagctgcagg tcgtgctcaa aggctcgcag
     2101 ggtccgctgc atggagctgg cactgggtct gggttcaggt ggtgccgtca ggcggggcag
     2161 cctcctccgc ttctcctcaa tctgcccttg aagctcccgg gcatcgctga agaacttatg
     2221 cagctcccga gaggcagcga gcagctgggc ccgggtgccc atgagttcca gcagctcagc
     2281 ccaggcctcg ttcagcccgt ccttccactc agccatggtg gccgctgctg tgtgaccaca
     2341 ctcaatcagc tcgtccacca tctggttgac cgccgccagc cgctcccgcc ctgcggttcc
     2401 tgtctcactg gcaaactctg agaatttctc ctgtagcacc gacacgtgtt cgaagtcctg
     2461 gcccagctct ggggagccag ctaccacctc cttctcggct atccagtgtt ccagctcatc
     2521 cacctggcgg ctgagctggt agagccagta ctgctgttcc aggctcaccc tgcgttcctc
     2581 gcccagctcc ttgagcgcca catagagacc ggtcgattgg aagtagaggt tctcgctagc
     2641 tttatcgtca tcgtctttgt agtccttgtc gtcgtcgtcc ttgtaatcct tatcatcatc
     2701 atctttatag tcgctagcca tggtaccgaa ttctttgcca aaatgatgag acagcacaat
     2761 aaccagcacg ttgcccagga gctgtaggaa aaagaagaag gcatgaacat ggttagcaga
     2821 ggctctagag ccgccggtca cacgccagaa gccgaacccc gccctgcccc gtcccccccg
     2881 aaggcagccg tccccccgcg gacagccccg aggctggaga gggagaaggg gacggcggcg
     2941 cggcgacgca cgaaggccct ccccgcccat ttccttcctg ccggcgccgc accgcttcgc
     3001 cccgcgcccg ctagaggggg tgcggcggcg cctcccagat ttcggctccg cacagatttg
     3061 ggacaaagga agtccctgcg ccctctcgca cgattaccat aaaaggcaat ggctgcggct
     3121 cgccgcgcct cgacagccgc cggcgctccg ggggccgccg cgcccctccc ccgagccctc
     3181 cccggcccga ggcggccccg ccccgcccgg cacccccacc tgccgccacc ccccgcccgg
     3241 cacggcgagc cccgcgccac gccccgtacg gagccccgca cccgaagccg ggccgtgctc
     3301 agcaactcgg ggaggggggt gcaggggggg ttgcagcccg accgacgcgc ccacaccccc
     3361 tgctcacccc cccacgcaca caccccgcac gcagcctttg ttcccctcgc agcccccccc
     3421 gcaccgcggg gcaccgcccc cggccgcgct cccctcgcgc acactgcgga gcgcacaaag
     3481 ccccgcgccg cgcccgcagc gctcacagcc gccgggcagc gcggagccgc acgcggcgct
     3541 ccccacgcac acacacacgc acgcaccccc cgagccgctc cccccgcaca aagggccctc
     3601 ccggagcccc tcaaggcttt cacgcagcca cagaaaagaa acaagccgtc attaaaccaa
     3661 gcgctaatta cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc
     3721 ggtcagtcag agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag
     3781 gcagcgcgca gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg
     3841 ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc
     3901 tctccgcctc gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc
     3961 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
     4021 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
     4081 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     4141 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     4201 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     4261 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     4321 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     4381 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     4441 tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6421 cccgcgcgtt ggccgattca ttaatgcagc ggatccagac atgataagat acattgatga
     6481 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6541 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttagct ttttgcaaaa
     6601 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6661 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6721 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6781 gagatgcgga tccgctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6841 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6901 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6961 catgattacg ccaagcttgg gctgcaggtc gagggatctc cataagagaa gagggacagc
     7021 tatgactggg agtagtcagg agaggaggaa aaatctggct agtaaaacat gtaaggaaaa
     7081 ttttagggat gttaaagaaa aaaataacac aaaacaaaat at
//
