LOCUS       sequence-413364-        8054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Shtn1 long isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8054)
  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 8054)
  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-SHTN1L
FEATURES             Location/Qualifiers
     source          1..8054
                     /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(3706..4280,4281..4283,4284..4292)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3873..3890)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4100..4119
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4293..4295,4296..4298,4299..5012)
                     /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(4293..4295,4296..4298,4299..5009)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4338..4359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4599..4618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4946..4967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5076..5095)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5141..5196
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5195..5214)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5557..5573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5557..5573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5570..5592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5581..5597
                     /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(5605..5611,5612..5629,5630..5635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5650..5671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5730..5925
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5776..5911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5931..6065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5968..5987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6022..6041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6133..6150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6304..6892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6384..6403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7063..7854,7855..7923))
                     /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     7686..7705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7924..8028)
                     /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 ccccgggggg gggggggggg gggggggggc
      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 aatcctgggg acacttaaca aatccactag ttcgagaagc ttaaaatccc ttggccctga
     3181 gaacagtgaa actgagttag agaggatttt acgtcgcaga aagctgacgg cagaagcaga
     3241 tagcagtagt cctaccggga tattagccac ctcagagtcc aagtccatgc cagtgttggg
     3301 ttctgtatcc agtgtaacaa aatcagcctt gaacaagaaa actctggagg cagaattcaa
     3361 caacccgtgt cccctaacac ctgagccagg tgaagggccc cgtaaattgg aaggatgcac
     3421 gaatcccaag gttacctttc agcctcccag taaaggtgga tacaggagaa aatgtgtggg
     3481 cagtgaaaac caagctgagc cagttgtagt attagatcct gtttccacac acgaacccca
     3541 aaccaaagac caggctgctg aaaaagaccc aactcaattc gaggaggagg gaggtgaaac
     3601 ccaaccagaa tacaaagaag acagcggtgg gaaaacagga gagacggaca gttccaactg
     3661 ctgatcgagg atatcgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc
     3721 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     3781 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     3841 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     3901 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     3961 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     4021 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     4081 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     4141 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     4201 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     4261 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca
     4321 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4381 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4441 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4501 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4561 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4621 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4681 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     4741 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     4801 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4861 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     4921 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4981 tcactctcgg catggacgag ctgtacaagt aaagcggccg gccgccagca cagtggtcga
     5041 tcgaccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     5101 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     5161 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     5221 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     5281 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     5341 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     5401 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     5461 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     5521 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5581 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5641 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5701 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     5761 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5821 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5881 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     5941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6061 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6121 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8041 aaaagtgcca cctg
//
