LOCUS       Exported                6689 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  IDOCKSalpha Donor Construct.
ACCESSION   .
VERSION     .
KEYWORDS    CMV-PKCa-mEGFP-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  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 6689)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6689
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      2629..2638
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2635..3354
                     /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             2635..3351
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2680..2701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2941..2960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3288..3309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3477..3598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3514..3533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3568..3587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3605..4060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3742..3763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3954..3973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4087..4191
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4193..4550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4216..4236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4401..4536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4463..4482
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4585..5379
                     /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(4639..4658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5249..5268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5567..5586)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5611..5658
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5987..6575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6476..6495
                     /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 ccgatggctg acgtcttccc ggccgccgag ccggtggcgc cgcaggacgt ggccaaccgc
      661 ttcgcccgca aaggggcgct gaggcagaag aacgtgcacg aggtgaagaa ccaccgcttc
      721 atcgcgcgct tcttcaagca gcccaccttc tgcagccact gcaccgactt catctggggg
      781 tttgggaaac aaggcttcca gtgccaagtt tgctgttttg tggttcacaa gaggtgccat
      841 gaatttgtta ctttttcttg tccgggggcg gataaaggac ccgacacaga tgacccgagg
      901 agcaagcaca agttcaagat ccacacgtat ggcagcccca ccttctgtga tcactgcggc
      961 tccctgctgt acggactcat ccaccagggg atgaaatgtg acacctgtga tatgaacgtg
     1021 cacaagcagt gcgtgatcaa cgtgcccagc ctctgcggga tggaccacac ggagaagagg
     1081 ggccgcatct acctgaaggc cgaggtcacg gatgaaaagt tgcacgtcac agtacgagac
     1141 gcgaaaaacc taatccctat ggatccaaat gggctttcag atccttacgt gaagctgaag
     1201 cttattcctg accccaagaa cgagagcaaa cagaaaacca agaccatccg ctcgacgctg
     1261 aacccccggt gggacgagtc cttcacgttc aaattaaaac cttctgataa agaccggcga
     1321 ctgtccgtgg aaatctggga ctgggatcga accacacgga acgacttcat ggggtccctt
     1381 tcctttgggg tctcggagct gatgaagatg ccggccagcg gatggtacaa gctgctgaac
     1441 caagaggagg gcgagtacta caacgtgccg atccccgaag gcgacgagga aggcaatgtg
     1501 gagctcaggc agaaattcga gaaagccaag cttggccctg ccggcaacaa agtcatcagt
     1561 ccctccgagg acaggagaca gccttccaac aacctgggca gagtgaagct cacggacttc
     1621 aacttcctca tggtgctggg caaaggcagc tttgggaagg tgatgctggc cgaccgaaag
     1681 gggacagagg agctgtacgc catcaagatc ctgaagaagg acgtggtcat ccaggacgac
     1741 gacgtggagt gcaccatggt ggagaagcgg gtcctggcgc tgctcgacaa gccgccgttc
     1801 ctgacgcagc tgcactcctg cttccagacg gtggaccggc tgtacttcgt catggagtac
     1861 gtcaacggcg gggacctcat gtaccacatc cagcaggtcg ggaagttcaa ggagccgcaa
     1921 gcagtgttct atgcagcaga gatttccatc gggctgttct ttcttcataa aagaggaatc
     1981 atttatcggg acctgaagtt agacaacgtc atgctggact cggaaggaca cattaagatc
     2041 gcggacttcg ggatgtgcaa ggagcacatg atggacggcg tcacgaccag gaccttctgc
     2101 gggacccccg actacatcgc cccagagata atcgcctatc agccgtacgg gaagtccgtg
     2161 gactggtggg cctacggcgt cctgttgtac gagatgttgg ccgggcagcc tccgttcgac
     2221 ggcgaggacg aggacgagct gttccagtcc atcatggagc acaacgtctc gtaccccaag
     2281 tccttgtcca aggaggccgt gtccatctgc aaagggctga tgaccaagca ccccgggaag
     2341 cggctgggct gcgggcccga gggcgagcgc gacgtgcggg agcatgcctt cttccggagg
     2401 atcgactggg agaagctgga gaaccgtgag atccagccac ccttcaagcc caaagtgtgc
     2461 ggcaaaggag cagagaactt tgacaagttc ttcacgcgag ggcagcctgt cttgacgccg
     2521 cccgaccagc tggtcatcgc taacatcgac cagtctgatt ttgaaggctt ctcctacgtc
     2581 aacccccagt tcgtgcaccc catcctgcag agcgcggtac caccggtcgc caccatggtg
     2641 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2701 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2761 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2821 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2881 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2941 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3001 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3061 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3121 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3181 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3241 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3301 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     3361 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3421 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3481 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3541 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3601 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3661 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3721 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3781 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3841 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3901 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3961 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4021 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4201 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4381 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4501 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4561 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4621 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4681 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4741 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4801 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4861 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4921 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4981 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5041 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5101 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5161 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5221 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5281 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5341 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5401 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5461 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5521 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5581 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5641 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5701 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5761 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5821 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6121 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6661 ctgtggataa ccgtattacc gccatgcat
//
