LOCUS       Exported                7380 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression construct driving the expression of a 
            fluorescent fusion protein consisting of EGFP linked to the 
            N-terminus of mouse neurofilament protein M.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-mNFM
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  AUTHORS   Yan Y, Jensen K, Brown A
  TITLE     The polypeptide composition of moving and stationary neurofilaments 
            in cultured sympathetic neurons.
  JOURNAL   Cell Motil Cytoskeleton. 2007 Apr;64(4):299-309.
  PUBMED    17285620
REFERENCE   2  (bases 1 to 7380)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7380
                     /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      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             613..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"
     misc_feature    1330..1395
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4168..4289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4205..4224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4259..4278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4296..4751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4433..4454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4645..4664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4778..4882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4884..5241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4907..4927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5092..5227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5154..5173
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5276..6070
                     /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(5330..5349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5940..5959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6258..6277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6302..6349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6678..7266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7167..7186
                     /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 caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccgg gatccgcctc caagatgagc tacacgctgg actcgctggg caacccgtcc
     1441 gcctaccggc gggtcaccga gacccgctcc agcttcagcc gcgtgagcgg ttccccgtcc
     1501 agcggcttcc gctcgcagtc ctggtcccgc ggctcgccca gcaccgtgtc ctcctcctac
     1561 aagcgcagcg cgctcgcccc gcgtctcgcc tacagctcgg ctatgctcag ctcggccgag
     1621 agcagcctcg acttcagcca gtcctcgtcg ctgctcaacg gcggctccgg cggcgactac
     1681 aaactgtccc gctctaacga gaaagagcag ctgcaggggc tgaacgaccg cttcgccggc
     1741 tacatcgaga aagtgcacta cttggaacaa cagaacaagg agatcgaagc agagatccag
     1801 gcactgcggc agaagcaggc ctcgcacgcc cagctgggtg atgcttacga ccaggagatc
     1861 cgagagctgc gcgccaccct cgagatggtg aaccacgaga aggctcaagt gcagctggac
     1921 tccgatcact tggaggaaga catccaccgg ctcaaggagc gcttcgagga ggaggcgcgg
     1981 ctgcgggacg acaccgaggc tgccattcgc gcgctgcgca aagacatcga agagtcgtcg
     2041 atggttaagg tggagctgga caagaaggtg cagtcgctgc aggatgaggt ggctttcctg
     2101 cggagcaatc acgaagagga ggtggccgac ctgctggctc agatccaggc gtcgcacatc
     2161 acggtagagc gcaaagatta cctgaagaca gacatctcca cggcgctgaa ggagatccgc
     2221 tcccagctcg agtgtcactc agaccagaac atgcaccagg ccgaagagtg gttcaaatgc
     2281 cgctacgcca agctcaccga ggcggccgag cagaacaagg aggccattcg ctctgccaag
     2341 gaagagatcg ccgagtaccg gcgccagctg cagtccaaga gcatcgagct cgagtcggtg
     2401 cgaggcacta aggagtccct ggagcggcag ctcagcgaca tcgaggagcg ccacaaccac
     2461 gacctcagca gctaccagga caccatccag cagttggaaa atgaacttcg gggaaccaag
     2521 tgggaaatgg ctcgtcattt gcgagaatac caggatctcc ttaacgtcaa gatggccctg
     2581 gacatcgaga tcgccgcgta caggaaactc ctagaggggg aagagaccag atttagcaca
     2641 ttttcaggaa gcatcaccgg gcctctgtac acacaccgac agccctcagt cacaatatcc
     2701 agtaagattc agaagaccaa agtcgaggcc cccaagctca aggtccaaca caaatttgtg
     2761 gaggagatca tcgaagaaac taaagtggaa gatgagaagt cagaaatgga agaaaccctc
     2821 acagccatcg cagaggagtt ggcagcctcc gccaaagagg agaaggaaga ggccgaagaa
     2881 aaggaggagg aaccagaagc cgaaaagtct cccgtgaagt ctcctgaggc taaggaagag
     2941 gaggaggaag gggaaaagga ggaagaagag gaaggccagg aggaagaaga ggaggaagat
     3001 gaaggtgtca agtcagacca ggcagaagag gggggatctg agaaggaagg ctccagtgaa
     3061 aaagatgaag gtgagcagga agaagaagaa ggagaaaccg aggcagaagg tgaaggagag
     3121 gaagcagaag ctaaggagga aaagaaaatt gagggaaagg ttgaggaagt ggctgtcaag
     3181 gaggaaatca aggtcgagaa gcctgagaaa gccaaatccc ctatgcccaa atcacccgtg
     3241 gaagaagtaa agccaaaacc agaggccaag gccgggaagg gtgagcacaa ggaggaagag
     3301 aaagttgagg aagagaagaa ggaagtcacc aaagaatcac ccaaggaaga gaaggtggag
     3361 aaaaaggagg agaagccaaa agatgttgca gataaaaaga aggccgagtc cccggtgaaa
     3421 gagaaggctg tggaggaggt gatcaccatc agcaagtcgg taaaggtgag cctagagaaa
     3481 gacaccaaag aggagaagct gcagccgcag gagaaggtga aggagaaggc agaggaggag
     3541 gggggcagtg aggaggaagg gagtgaccgt agcccgcagg agtccaagaa ggaagacata
     3601 gctatcaatg gggaggtgga aggaaaagag gaggaggagc aggaaactca ggagaagggc
     3661 agtgggcggg aggaggagaa aggggtggtc actaatggct tagatgtgag ccctgcagag
     3721 gagaagaaag gagaggatag cagtgatgat aaagtggtgg tcaccaagaa ggtagaaaag
     3781 atcaccagcg aggggggcga tggtgctacc aaatacatca ccaaatctgt aaccgtcact
     3841 caaaaggttg aagagcatga ggagaccttt gaggagaagc tggtctcaac taaaaaggta
     3901 gaaaaggtca cttcacacgc catagtcaag gaagtcaccc agggtgacta agatcggagt
     3961 ccgttgcaaa aggttaagcc atacgacaat ttcaaaatgc atgtgattga cagcttcaaa
     4021 acagaatggg ttctcccagg atccaccgga tctagataac tgatcataat cagccatacc
     4081 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4141 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4201 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4261 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4321 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4381 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4441 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4501 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4561 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4621 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4681 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4741 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4861 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4921 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4981 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5041 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5101 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5161 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5221 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5281 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5341 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5401 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5461 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5521 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5581 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5641 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5701 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5761 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5821 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5881 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5941 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6001 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6061 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6121 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6181 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6241 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6301 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6361 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6421 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6481 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6541 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6601 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     6841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7021 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7081 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7141 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7201 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7261 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7321 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
