LOCUS       sequence-413486-        6064 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 6064)
  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 6064)
  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: pEGFP-C1-SHTN1S
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2852..2973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2889..2908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2943..2962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2980..3435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3117..3138)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3329..3348
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3462..3566
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3568..3925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3568..3759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3591..3611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3776..3911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3960..4754
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3960..4754
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4014..4033)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4624..4643
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4942..4961)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4986..5033
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5362..5950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5851..5870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct aacagctcgg acgaggagaa gcagctgcag
     1381 ctcatcacca gcttgaagga gcaagccatt ggcgaatatg aagaccttcg agcagagaac
     1441 cagaaaacaa aggagaagtg tgacaaaatt aggcaagaac gagatgaagc tgttaaaaaa
     1501 ctggaagagt ttcagaaaat ttcacatatg gttatagagg aggtgaattt catgcagaac
     1561 catcttgaaa tagagaagac atgtcgagag agcgctgaag ccttggcgac aaagctaaat
     1621 aaagaaaata aaacactgaa aagaatcagc atgctataca tggccaaact ggggccagac
     1681 gtaattacag aggagataaa cattgatgat gatgacccag ccaccgacac agatgctgcg
     1741 gctgagacct gtgtgtctgt tcagtgtcag aagcaaatca aagaacttcg agatcaaatt
     1801 gtgtctgttc aggaagaaaa gaaggtgtta gccatcgagc tggaaaacct caagagcaaa
     1861 ctcggggagg tgatggagga ggtaaataaa gttaagcaag aaaaagctgt tttaaattca
     1921 gaagtccttg agcagaggaa agtcttagaa aaatgcaaca gagtgtccat gttggctgtt
     1981 gaagagtatg aggaactgca agtgaacctg gaactggaga aggaccttcg caagaaagca
     2041 gagtcttttg cacaagagat gttcattgaa caaaacaaac tgaagagaca aagccacctt
     2101 ctgctgcaga gctcccttcc tgaccagcag cttttgaaag ctttagacga aaacgcaaaa
     2161 cttatccagc agcttgaaga agagaggatc cagcatcaga aaaaggtcaa agagctggag
     2221 gagcggctgg agaatgaagc acttcacaaa gagatccata acctcagaca acagctggag
     2281 cttctggaag acgacaagag ggagctagag cagaaatacc agagctcgga ggagaaggcc
     2341 cggaacctga agcattcagt ggatgaactt cagaagcgag tgaaccagtc tgagaattcg
     2401 gtacctcccc cgcctcctcc tcctccacct ctcccccctc cacctcccaa tccaatccgg
     2461 tccctcatgt ctatgatccg gaagcgatct caccccagtg gcaatagtgc taagaaagaa
     2521 aagacaactc agccagagac agctgaggaa gtcacagacc tgaagaggca agcagtggaa
     2581 gagatgatgg acagaattaa gaagggagtt catcttagac cggttaacca gacagccaga
     2641 cccaaggcaa agccagactc tctcaagggc tcagaaagtg cggtggatga gctgaaggga
     2701 atcctggcct cccagtaacc cgggatccac cggatctaga taactgatca taatcagcca
     2761 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2821 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2881 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2941 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     3001 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3061 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3121 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3181 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3241 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3301 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3361 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3421 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3481 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3541 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3841 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3901 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3961 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4021 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4081 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4141 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4201 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4261 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4321 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4381 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4441 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4501 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4561 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4621 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4681 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4741 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4801 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4861 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4921 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4981 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5041 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5101 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5161 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5221 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5281 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5341 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5401 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5461 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5521 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5581 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5641 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5701 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5761 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5821 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5881 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5941 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6001 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6061 gcat
//
