LOCUS       sequence-422467-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of bII spectrin 
            (repeats 13-15) in NIH/3T3 cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  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 7114)
  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-bII spectrin (repeats 13-15)
FEATURES             Location/Qualifiers
     source          1..7114
                     /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(2608..2628)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2635..2706)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2749..2768)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2776..3792
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2830..2852)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3793..4069)
                     /label=chicken beta-actin promoter
     enhancer        4071..4450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4481..4585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4586..4654,4655..5446)
                     /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(4804..4823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5617..6205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6106..6125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6359..6376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6444..6578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6468..6487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6522..6541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6584..6779)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6598..6733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6838..6859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6874..6879,6880..6897,6898..6904)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6912..6928
                     /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     6936..6952
                     /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 acttaagaat aagcttctcc acttcgtcta cactctggcc
     1501 aatttcacgg ctggataggt atggttcctg tccaagcagc caagcctctg ccacactggc
     1561 atcccttgag aactgatgga cctccaaaat cagtcttaac cactcccacc ggtcttccca
     1621 cttgtcaatc atttcttttc ttttctctgt cagctgcagt aacttttcct tgatctcctc
     1681 agaagcatag tgtttccgtg ccagcaggga tttcccaagc tcaatgcagg ctgtaaagct
     1741 gtcattacga gcatcaattt cagctttgat accttgatga ttattcatta acagttcaac
     1801 agatgacaca tcccgtggtt tctcctgggc ctcgatctgc cggatgacat cttccatcca
     1861 gagcatgagg tcacgcacca tgctgaagaa gcggaacttg tctcctgtgt ctaccagccg
     1921 caccctgcga ccctcacaag catccagcag ggacttccag gcttccagga cctcattctc
     1981 acgcttctgg atgtcatcag ccttgtcccc tgcataggct gcctggaggc gagctgcatc
     2041 ctcctgcagc tgcctcacct gagtgcccag agcttggatg tcgtgctcaa aggtggtgtg
     2101 cattctctgt aaagtttcca cagtgttttg atctcttcca agctcctcag ggagtttctt
     2161 gtgtttgtcc tggattcggc caaagatctc cttggcatca tggtaaaact tatgaagttc
     2221 atatgaggca gcaagaatct gtgttcttgt gtcaatgagc tccaggaggt cagcccaggc
     2281 ttcattgaga ccatctttcc actcagcaat ggtggcagca tctgaatgtc cagagttgat
     2341 gagttcatct gccatgttat taactgtatc cacacgctcc tgcccaatgt ttcctgtgtc
     2401 tcgagcaaat tctcggaacc gttcttgtaa catcgtgaca tgctcatagt cctgtcccaa
     2461 ctcatgggag cctgcgacca cttccctctc agcgatccac tgttccaggt catccacctc
     2521 tctgttgagc tggaacagcc tgtgcctctc atcaagtttt cctctcctct cctcagcaag
     2581 gtccttcagg ccagcataca gaccggtcga ttggaagtag aggttctcgc tagctttatc
     2641 gtcatcgtct ttgtagtcct tgtcgtcgtc gtccttgtaa tccttatcat catcatcttt
     2701 atagtcgcta gccatggtac cgaattcttt gccaaaatga tgagacagca caataaccag
     2761 cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag cagaggctct
     2821 agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc cccgaaggca
     2881 gccgtccccc cgcggacagc cccgaggctg gagagggaga aggggacggc ggcgcggcga
     2941 cgcacgaagg ccctccccgc ccatttcctt cctgccggcg ccgcaccgct tcgccccgcg
     3001 cccgctagag ggggtgcggc ggcgcctccc agatttcggc tccgcacaga tttgggacaa
     3061 aggaagtccc tgcgccctct cgcacgatta ccataaaagg caatggctgc ggctcgccgc
     3121 gcctcgacag ccgccggcgc tccgggggcc gccgcgcccc tcccccgagc cctccccggc
     3181 ccgaggcggc cccgccccgc ccggcacccc cacctgccgc caccccccgc ccggcacggc
     3241 gagccccgcg ccacgccccg tacggagccc cgcacccgaa gccgggccgt gctcagcaac
     3301 tcggggaggg gggtgcaggg ggggttgcag cccgaccgac gcgcccacac cccctgctca
     3361 cccccccacg cacacacccc gcacgcagcc tttgttcccc tcgcagcccc ccccgcaccg
     3421 cggggcaccg cccccggccg cgctcccctc gcgcacactg cggagcgcac aaagccccgc
     3481 gccgcgcccg cagcgctcac agccgccggg cagcgcggag ccgcacgcgg cgctccccac
     3541 gcacacacac acgcacgcac cccccgagcc gctccccccg cacaaagggc cctcccggag
     3601 cccctcaagg ctttcacgca gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta
     3661 attacagccc ggaggagaag ggccgtcccg cccgctcacc tgtgggagta acgcggtcag
     3721 tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg
     3781 cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct
     3841 cgccataaaa ggaaactttc ggagcgcgcc gctctgattg gctgccgccg cacctctccg
     3901 cctcgccccg ccccgcccct cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc
     3961 cccgccccca tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg
     4021 gagggggggg agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata
     4081 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     4141 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     4201 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     4261 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     4321 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     4381 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     4441 aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4501 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4561 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4621 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4681 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4741 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4801 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4861 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4921 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4981 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5041 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5101 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5161 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5221 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5281 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5341 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5401 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5461 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5521 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5581 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5641 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5701 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5761 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5821 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5881 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5941 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6001 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6061 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6121 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6181 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6241 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6301 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6361 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6421 ttggccgatt cattaatgca gcggatccag acatgataag atacattgat gagtttggac
     6481 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6541 ctttatttgt aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc
     6601 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     6661 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     6721 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg
     6781 gatccgctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6841 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6901 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6961 cgccaagctt gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg
     7021 ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg
     7081 atgttaaaga aaaaaataac acaaaacaaa atat
//
