LOCUS       sequence-223040-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IDOCKSgamma Donor Construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  AUTHORS   Colgan LA, Hu M, Misler JA, Parra-Bueno P, Moran CM, Leitges M, 
            Yasuda R
  TITLE     PKCalpha integrates spatiotemporally distinct Ca(2+) and autocrine 
            BDNF signaling to facilitate synaptic plasticity.
  JOURNAL   Nat Neurosci. 2018 Aug;21(8):1027-1037. doi: 
            10.1038/s41593-018-0184-3. Epub 2018 Jul 16.
  PUBMED    30013171
REFERENCE   2  (bases 1 to 6764)
  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: 
            "10.1038/s41593-018-0184-3"; journalName: "Nat Neurosci"; date: 
            "2018-08"; volume: "21"; issue: "8"; pages: "1027-1037"
COMMENT     Sequence Label: CMV-PKCg-mEGFP-N1
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..568
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     regulatory      2704..2713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2710..2712,2713..2715,2716..3429)
                     /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(2710..2712,2713..2715,2716..3426)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2755..2776)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3016..3035)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3363..3384
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3552..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3589..3608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3643..3662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3680..4135)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3817..3838)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4029..4048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4162..4266
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4268..4625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4268..4459
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4291..4311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4660..5454
                     /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             4660..5454
                     /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(4714..4733)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5324..5343
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5642..5661)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5686..5733
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6062..6650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6551..6570
                     /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 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      481 atataagcas rgggatttcc aagtctccac cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccgatggcgg gtctgggccc tggcggaggc gactcagagg ggggaccccg acccctgttt
      661 tgcagaaagg gggcgctgag gcagaaggtg gtccacgagg tgaagagcca caagttcacc
      721 gctcgtttct tcaagcagcc aaccttctgc agtcactgta ccgacttcat ctggggcatt
      781 ggaaagcagg gcctgcaatg tcaagtctgt agctttgtgg ttcaccgccg atgccacgaa
      841 tttgtgacct tcgagtgtcc aggcgctgga aagggccccc agacggacga ccctcgcaac
      901 aagcacaagt tccgtctgca cagctacagc agtcccacct tctgcgacca ctgtggctcc
      961 ctcctttacg ggctggtgca ccagggcatg aaatgttcct gttgcgagat gaatgtgcac
     1021 cggcgctgtg tgcgcagcgt gccctccctt tgcggtgtgg accacacaga gcgccgtgga
     1081 cgtctgcaac tggaaatccg ggctcctacg tcggatgaga tccatattac tgttggcgag
     1141 gcccggaacc tcattcctat ggaccccaat ggcctgtctg atccctacgt gaaactgaag
     1201 ctgatcccgg accctcggaa cctgacaaaa cagaagacaa agaccgtgaa agccacactg
     1261 aatcccgtgt ggaacgagac cttcgtgttc aacctgaagc caggggatgt agagcgccgg
     1321 ctcagtgtgg aggtgtggga ttgggatagg acatcccgaa atgacttcat gggtgccatg
     1381 tcctttggtg tctcagagct actcaaggcc cctgtggatg gatggtacaa gttactgaac
     1441 caggaggagg gcgaatatta caatgtaccg gtggccgatg ctgacaactg cagcctcctc
     1501 cagaagtttg aggcctgcaa ttaccccttg gaattgtatg agagagtgcg gatgggcccc
     1561 tcttcctccc ccatcccttc tccatccccc agtcctacgg actccaagcg atgcttcttt
     1621 ggtgccagcc caggacgcct gcatatctct gacttcagct tcctcatggt tctagggaaa
     1681 ggcagttttg ggaaggtaat gctggcagag cgcagaggct ccgacgaact ctatgccatc
     1741 aagatactga agaaagacgt catcgtccag gatgacgatg tagactgcac cctcgtagag
     1801 aagcgtgtcc tggcattggg aggccgaggt cctggaggcc ggccacactt tctcacgcag
     1861 cttcactcca cctttcagac tccggaccgc ctgtattttg tgatggagta tgtcactggg
     1921 ggcgatttaa tgtaccacat ccagcaactg ggcaagttta aggagcctca tgcagcattc
     1981 tacgctgcgg aaatcgccat aggcctcttc ttccttcaca accagggcat catctacagg
     2041 gacctcaagt tggataatgt gatgctggat gctgaaggac acatcaagat cacagacttt
     2101 ggcatgtgta aagagaatgt cttccctggg tccacaaccc gcaccttctg tggcacccca
     2161 gactacatag cacctgagat cattgcctat cagccctacg ggaagtctgt cgactggtgg
     2221 tcttttgggg tcctgctgta tgagatgttg gcaggacagc caccctttga tggggaagat
     2281 gaggaggagt tgtttcaagc catcatggaa caaactgtca cctatcccaa gtcactttcc
     2341 cgggaagctg tggccatctg caaagggttc ctgaccaagc acccaggaaa acgcctgggc
     2401 tcagggccag atggggaacc caccatccgg gctcatggct ttttccgttg gatcgattgg
     2461 gagaggttgg agagactgga aattgcacct cctttcagac cacgtccgtg tggccgcagt
     2521 ggcgaaaact ttgacaagtt cttcacgcgg gcagcgccag cactgacccc gccagaccgc
     2581 ttggttctag ccagcatcga ccaagctgat ttccagggct ttacttatgt gaacccggac
     2641 ttcgtgcacc cagatgcccg cagccccaca agccctgtgc ccgtgcctgt catgccaccg
     2701 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2761 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2821 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2881 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2941 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3001 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3061 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3121 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3181 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3241 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3301 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3361 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3421 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     3481 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3541 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3661 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3721 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3781 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3841 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3901 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3961 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4021 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4081 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4201 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4261 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4681 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4741 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4801 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4861 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4921 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4981 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5041 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5101 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5161 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5221 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5281 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5341 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5401 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5461 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5521 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5581 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5641 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5701 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5761 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5821 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5881 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6181 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6721 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
