LOCUS       sequence-413367-        7529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Shtn1 short isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7529)
  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 7529)
  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-SHTN1S
FEATURES             Location/Qualifiers
     source          1..7529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1739..1748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1748..1771
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3181..3755,3756..3758,3759..3767)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3348..3365)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3575..3594
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3768..3770,3771..3773,3774..4487)
                     /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(3768..3770,3771..3773,3774..4484)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3813..3834)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4074..4093)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4421..4442
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4551..4570)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4616..4671
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4670..4689)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5032..5048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5032..5048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5045..5067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5056..5072
                     /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(5080..5086,5087..5104,5105..5110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5125..5146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5205..5400
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5251..5386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5406..5540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5443..5462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5497..5516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5608..5625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5779..6367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5859..5878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6538..7329,7330..7398))
                     /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     7161..7180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7399..7503)
                     /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 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagc
     1741 caccatggac tacaaggacg acgacgacaa gaacagctcg gacgaggaga agcagctgca
     1801 gctcatcacc agcttgaagg agcaagccat tggcgaatat gaagaccttc gagcagagaa
     1861 ccagaaaaca aaggagaagt gtgacaaaat taggcaagaa cgagatgaag ctgttaaaaa
     1921 actggaagag tttcagaaaa tttcacatat ggttatagag gaggtgaatt tcatgcagaa
     1981 ccatcttgaa atagagaaga catgtcgaga gagcgctgaa gccttggcga caaagctaaa
     2041 taaagaaaat aaaacactga aaagaatcag catgctatac atggccaaac tggggccaga
     2101 cgtaattaca gaggagataa acattgatga tgatgaccca gccaccgaca cagatgctgc
     2161 ggctgagacc tgtgtgtctg ttcagtgtca gaagcaaatc aaagaacttc gagatcaaat
     2221 tgtgtctgtt caggaagaaa agaaggtgtt agccatcgag ctggaaaacc tcaagagcaa
     2281 actcggggag gtgatggagg aggtaaataa agttaagcaa gaaaaagctg ttttaaattc
     2341 agaagtcctt gagcagagga aagtcttaga aaaatgcaac agagtgtcca tgttggctgt
     2401 tgaagagtat gaggaactgc aagtgaacct ggaactggag aaggaccttc gcaagaaagc
     2461 agagtctttt gcacaagaga tgttcattga acaaaacaaa ctgaagagac aaagccacct
     2521 tctgctgcag agctcccttc ctgaccagca gcttttgaaa gctttagacg aaaacgcaaa
     2581 acttatccag cagcttgaag aagagaggat ccagcatcag aaaaaggtca aagagctgga
     2641 ggagcggctg gagaatgaag cacttcacaa agagatccat aacctcagac aacagctgga
     2701 gcttctggaa gacgacaaga gggagctaga gcagaaatac cagagctcgg aggagaaggc
     2761 ccggaacctg aagcattcag tggatgaact tcagaagcga gtgaaccagt ctgagaattc
     2821 ggtacctccc ccgcctcctc ctcctccacc tctcccccct ccacctccca atccaatccg
     2881 gtccctcatg tctatgatcc ggaagcgatc tcaccccagt ggcaatagtg ctaagaaaga
     2941 aaagacaact cagccagaga cagctgagga agtcacagac ctgaagaggc aagcagtgga
     3001 agagatgatg gacagaatta agaagggagt tcatcttaga ccggttaacc agacagccag
     3061 acccaaggca aagccagact ctctcaaggg ctcagaaagt gcggtggatg agctgaaggg
     3121 aatcctggcc tcccagtaat cgaggatatc gcggccgcta cgtaaattcc gccccccccc
     3181 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     3241 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     3301 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     3361 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     3421 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     3481 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     3541 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     3601 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     3661 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     3721 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     3781 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3841 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3901 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3961 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     4021 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4081 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4141 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4201 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     4261 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     4321 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     4381 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4441 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccggccgc
     4501 cagcacagtg gtcgatcgac caatgccctg gctcacaaat accactgaga tctttttccc
     4561 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4621 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4681 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4741 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     4801 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     4861 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     4921 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4981 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     5041 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5101 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5161 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     5221 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5281 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5341 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5401 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5461 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5521 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7501 gcgcacattt ccccgaaaag tgccacctg
//
