LOCUS       Exported                6737 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  ITRACKbeta/IDOCKSbeta Donor construct.
ACCESSION   .
VERSION     .
KEYWORDS    CMV-PKCb1-mEGFP-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6737
                     /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"
     CDS             2686..3402
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2728..2749)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2989..3008)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3336..3357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3525..3646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3562..3581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3616..3635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3653..4108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4002..4021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4135..4239
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4241..4598
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4264..4284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4449..4584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4511..4530
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4633..5427
                     /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(4687..4706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5297..5316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5615..5634)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5659..5706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6035..6623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6524..6543
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcggatggct
      661 gacccggctg cggggccgcc gccgagcgag ggcgaggaga gcacggtgcg cttcgcccgc
      721 aaaggcgccc tccggcagaa gaacgtgcac gaggtgaaga accacaaatt caccgcccgc
      781 ttcttcaagc agcccacctt ctgcagccac tgcaccgact tcatttgggg cttcgggaaa
      841 cagggattcc agtgtcaagt ctgctgcttt gttgtacaca agcgctgcca tgaattcgtc
      901 acgttctcct gccctggtgc agacaagggc ccggcctctg atgacccacg gagcaaacac
      961 aagtttaaga tccacaccta ctccagccct accttctgtg accactgtgg atcactgctg
     1021 tatgggctca tccaccaggg gatgaaatgc gacacctgta tgatgaatgt ccacaagcgc
     1081 tgcgtgatga acgtccccag cctctgtggc accgaccaca cagaacgccg tggccgcatc
     1141 tacatccagg cccacatcga cagggaggtc ctcatcgttg ttgtaagaga tgctaaaaat
     1201 ctggtaccta tggaccccaa cggcttgtca gatccctacg taaaactgaa actgatccct
     1261 gatcccaaaa gtgagagcaa gcagaagacc aagactatca aatgctccct caacccggag
     1321 tggaacgaaa ccttcagatt tcagctgaag gaatcagaca aagacagaag actgtccgta
     1381 gagatctggg attgggacct gaccagcagg aatgacttca tgggatctct gtcgtttggg
     1441 atttcagaac tacagaaagc cggagtggat ggctggttca agttactaag ccaggaagaa
     1501 ggcgagtact ttaatgtgcc ggtgccgccg gaaggaagcg agggcaatga agagctgcgg
     1561 cagaagtttg agagagccaa gattggccaa ggtaccaagg ctccagaaga aaagacagcg
     1621 aacactatat ccaaatttga caacaatggc aacagggacc ggatgaaact gaccgatttt
     1681 aacttcctga tggtgctggg gaaaggcagc tttggcaagg tcatgctctc agagcggaag
     1741 ggtacagatg aactctatgc cgtgaagatc ctgaagaaag atgtggtgat ccaagatgac
     1801 gatgtggagt gcacaatggt ggagaagagg gtgctggccc tgcctgggaa gcccccattc
     1861 ctgactcagc tccattcctg cttccagacc atggaccgcc tctactttgt gatggagtat
     1921 gtgaacgggg gcgacctcat gtaccacatc caacaagttg gccgtttcaa ggagccccat
     1981 gctgtatttt acgctgcaga gattgccatc ggtcttttct tcttgcagag caagggcatc
     2041 atttaccgtg acctgaaact tgacaacgtg atgctggatt ccgaggggca catcaaaatc
     2101 gctgactttg gcatgtgtaa agagaatatc tgggatgggg tgacaaccaa gacattctgt
     2161 ggcactccag actacattgc cccagagatc attgcttatc agccctacgg gaagtctgtg
     2221 gactggtggg cgtttggagt cctgctgtat gaaatgttgg ctggccaggc accttttgaa
     2281 ggggaggatg aggatgaact cttccagtca atcatggagc acaacgtggc gtatcccaag
     2341 tccatgtcta aggaagctgt ggcaatctgc aaagggctaa tgaccaaaca cccaggcaag
     2401 cgcctgggtt gtgggcctga aggggaacga gacattaagg agcatgcatt tttccggtat
     2461 atcgactggg agaaactcga acgcaaggag attcagccac cttataaacc aaaagcttgt
     2521 gggcgaaacg ctgaaaactt cgaccggttt ttcacccgcc atccaccagt cctaacacct
     2581 cctgaccagg aagtcatcag gaatattgac caatcagaat tcgaaggatt ttcctttgtt
     2641 aactctgaat ttttaaaacc cgaagtcaag agcctcgagg cactggtgag caagggcgag
     2701 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2761 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2821 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2881 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2941 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3001 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3061 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3121 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3181 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3241 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3301 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3361 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     3421 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3481 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3541 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3601 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3661 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4081 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4141 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4201 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4261 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4321 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4441 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4501 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4561 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4801 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5161 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5401 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5461 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5581 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5641 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5701 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5761 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5821 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6721 gtattaccgc catgcat
//
