LOCUS       sequence-413362-        8055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Shtn1 long isoform with RG mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Zhang M, Ergin V, Lin L, Stork C, Chen L, Zheng S
  TITLE     Axonogenesis Is Coordinated by Neuron-Specific Alternative Splicing 
            Programming and Splicing Regulator PTBP2.
  JOURNAL   Neuron. 2019 Feb 20;101(4):690-706.e10. doi: 
            10.1016/j.neuron.2019.01.022. Epub 2019 Feb 4.
  PUBMED    30733148
REFERENCE   2  (bases 1 to 8055)
  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.2019.01.022"; journalName: "Neuron"; date: 
            "2019-02-20- 20"; volume: "101"; issue: "4"; pages: "690-706.e10"
COMMENT     Sequence Label: pCAGIG-FLAG-SHTN1LRRR>GGG
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1740..1749
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1749..1772
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3707..4281,4282..4284,4285..4293)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3874..3891)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4101..4120
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4294..4296,4297..4299,4300..5013)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4294..4296,4297..4299,4300..5010)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4339..4360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4600..4619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4947..4968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5077..5096)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5142..5197
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5196..5215)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5558..5574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5558..5574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5582..5598
                     /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)."
     promoter        complement(join(5606..5612,5613..5630,5631..5636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5651..5672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5731..5926
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5777..5912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5932..6066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5969..5988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6023..6042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6134..6151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6305..6893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6385..6404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7064..7855,7856..7924))
                     /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     7687..7706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7925..8029)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgag
     1741 ccaccatgga ctacaaggac gacgacgaca agaacagctc ggacgaggag aagcagctgc
     1801 agctcatcac cagcttgaag gagcaagcca ttggcgaata tgaagacctt cgagcagaga
     1861 accagaaaac aaaggagaag tgtgacaaaa ttaggcaaga acgagatgaa gctgttaaaa
     1921 aactggaaga gtttcagaaa atttcacata tggttataga ggaggtgaat ttcatgcaga
     1981 accatcttga aatagagaag acatgtcgag agagcgctga agccttggcg acaaagctaa
     2041 ataaagaaaa taaaacactg aaaagaatca gcatgctata catggccaaa ctggggccag
     2101 acgtaattac agaggagata aacattgatg atgatgaccc agccaccgac acagatgctg
     2161 cggctgagac ctgtgtgtct gttcagtgtc agaagcaaat caaagaactt cgagatcaaa
     2221 ttgtgtctgt tcaggaagaa aagaaggtgt tagccatcga gctggaaaac ctcaagagca
     2281 aactcgggga ggtgatggag gaggtaaata aagttaagca agaaaaagct gttttaaatt
     2341 cagaagtcct tgagcagagg aaagtcttag aaaaatgcaa cagagtgtcc atgttggctg
     2401 ttgaagagta tgaggaactg caagtgaacc tggaactgga gaaggacctt cgcaagaaag
     2461 cagagtcttt tgcacaagag atgttcattg aacaaaacaa actgaagaga caaagccacc
     2521 ttctgctgca gagctccctt cctgaccagc agcttttgaa agctttagac gaaaacgcaa
     2581 aacttatcca gcagcttgaa gaagagagga tccagcatca gaaaaaggtc aaagagctgg
     2641 aggagcggct ggagaatgaa gcacttcaca aagagatcca taacctcaga caacagctgg
     2701 agcttctgga agacgacaag agggagctag agcagaaata ccagagctcg gaggagaagg
     2761 cccggaacct gaagcattca gtggatgaac ttcagaagcg agtgaaccag tctgagaatt
     2821 cggtacctcc cccgcctcct cctcctccac ctctcccccc tccacctccc aatccaatcc
     2881 ggtccctcat gtctatgatc cggaagcgat ctcaccccag tggcaatagt gctaagaaag
     2941 aaaagacaac tcagccagag acagctgagg aagtcacaga cctgaagagg caagcagtgg
     3001 aagagatgat ggacagaatt aagaagggag ttcatcttag accggttaac cagacagcca
     3061 gacccaaggc aaagccagac tctctcaagg gctcagaaag tgcggtggat gagctgaagg
     3121 gaatcctggg gacacttaac aaatccacta gttcgagaag cttaaaatcc cttggccctg
     3181 agaacagtga aactgagtta gagaggattt taggaggtgg caagctgacg gcagaagcag
     3241 atagcagtag tcctaccggg atattagcca cctcagagtc caagtccatg ccagtgttgg
     3301 gttctgtatc cagtgtaaca aaatcagcct tgaacaagaa aactctggag gcagaattca
     3361 acaacccgtg tcccctaaca cctgagccag gtgaagggcc ccgtaaattg gaaggatgca
     3421 cgaatcccaa ggttaccttt cagcctccca gtaaaggtgg atacaggaga aaatgtgtgg
     3481 gcagtgaaaa ccaagctgag ccagttgtag tattagatcc tgtttccaca cacgaacccc
     3541 aaaccaaaga ccaggctgct gaaaaagacc caactcaatt cgaggaggag ggaggtgaaa
     3601 cccaaccaga atacaaagaa gacagcggtg ggaaaacagg agagacggac agttccaact
     3661 gctgatcgag gatatcgcgg ccgctacgta aattccgccc cccccccccc tctccctccc
     3721 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     3781 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     3841 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     3901 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     3961 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4021 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4081 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     4141 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     4201 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     4261 tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc
     4321 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4381 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4441 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4501 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4561 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     4621 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     4681 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     4741 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     4801 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     4861 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     4921 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4981 atcactctcg gcatggacga gctgtacaag taaagcggcc ggccgccagc acagtggtcg
     5041 atcgaccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     5101 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     5161 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     5221 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     5281 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     5341 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5401 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5461 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     5521 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5581 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5701 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5881 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     5941 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6001 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6061 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6121 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6181 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6241 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6301 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6361 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6421 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6481 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6541 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6601 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6661 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6721 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6781 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6841 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6961 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7021 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7081 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7141 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7201 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7261 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7321 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7381 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7441 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7501 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7561 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7621 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7681 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7741 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7801 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7861 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7921 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8041 gaaaagtgcc acctg
//
