LOCUS       sequence-268763-        5327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron-specific expression of membrane targeted EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5327)
  AUTHORS   Santos TE, Schaffran B, Broguiere N, Meyn L, Zenobi-Wong M, Bradke F
  TITLE     Axon Growth of CNS Neurons in Three Dimensions Is Amoeboid and 
            Independent of Adhesions.
  JOURNAL   Cell Rep. 2020 Jul 21;32(3):107907. doi: 
            10.1016/j.celrep.2020.107907.
  PUBMED    32698008
REFERENCE   2  (bases 1 to 5327)
  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"; journalName: "Cell Rep.";
            date: "2020-07-21"; volume: "32(3):107907. doi"; pages: " 
            10.1016/j.celrep.2020.107907"
COMMENT     Sequence Label: pTub-alpha-1-mtGFP
FEATURES             Location/Qualifiers
     source          1..5327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..198
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     438..459
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     rep_origin      complement(1875..2463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1955..1974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2792..2839
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2864..2883
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3071..3865)
                     /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             complement(3071..3865)
                     /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(3182..3201)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3792..3811
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3900..4257)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3914..4049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4066..4257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4214..4234
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4259..4363)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4390..4845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4477..4496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4687..4708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4852..4973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4863..4882)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4917..4936
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5112..5327,1..498,499..501,502..504))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5157..5178)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
       61 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      121 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      181 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      241 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      301 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      361 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
      421 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      481 cagctcctcg cccttgctca ccatgatctt ttggtcctca tcattctttt caacctgttt
      541 ggttcttctc atacagcaca gcatggtggc gaccggtgga tcatcctcga ctgcagaatt
      601 cgaagcttga gctcgagatc tgagtccggt agcgctagtg gatcccccgc tcaccggtgt
      661 tgctgcttcg cggctgccgg gtggatggcg gaggcgagga ggaggtgttg cttcttacag
      721 cgcgactctt aagcggtcga tgtaagagaa cctgcggcac ttgggcggat gcgcctcctt
      781 atatacagct cggtcaccat ggggatgggc caaggccttt tcctgttggt ccagacccct
      841 ccgtctgccc agagaagccg cgacaggaga gtggtgattg gtgcaagcga catggtatga
      901 ggtaatctct ccccccttcc ccccctcgct agcatccgta gattttgcat ccttcagaca
      961 aaggcctcgc aaaagcaact agccgagcat ccattatggg gtggagctgg ggaggggtag
     1021 agagggaaga aagaaaacgg gggttgggtg attatacgga gcttaaacct taaactgtag
     1081 ttaggataat ttaaggtttt ccccaacact taatcacaga atttgtgttg agattcttta
     1141 tggtaaattc cagcttttca tttatactac actaaattta aatacgttcc gtcaggattc
     1201 aacatctctg ttcgctttaa ataaagtagc aagagctatg ggaatgatgg tcaaaaaggc
     1261 gtgggtaaat gtttagtttt acagttgtat tattgtggtt attattatta ctattattat
     1321 taatatttga gcaaaattca gaagctcagt atgaatagac cacgcctatg accctcaaca
     1381 gcacccggga ggcagaggca ggaagacttg cagctcaagg tcatcataac cgcccttagt
     1441 ggtttatgcc tgtaaatccc agcattggga agcggaggac tgcggttatt ttgtacccgg
     1501 cctggatata tagtgtgata tcctgtctca aaaaccaaaa acaaacaaag agagagagag
     1561 agagaaagga aaaaagagaa acaatataca aagtagtact taacgcttta ctagagagct
     1621 tgctttcttt agcggtagtg ctctcccatc acccctcccc gctcgccccc ttacccgacc
     1681 ctacctcacc ccagagcctg tgttattttg tccctagaaa tgagccatcc ctcctaatac
     1741 ggaattcagc ataataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     1801 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1921 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2281 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2341 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2401 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2461 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2521 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     2641 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     2701 gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     2761 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     2821 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     2881 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     2941 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     3001 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     3061 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3121 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3181 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3241 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3301 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     3361 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     3421 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3481 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3541 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3601 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3661 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     3721 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3781 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3841 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     3901 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     3961 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     4021 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     4081 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     4141 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     4201 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
     4261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4381 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4441 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4501 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4561 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4621 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4681 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4741 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4801 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcc ttaagataca
     4861 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4921 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4981 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5041 agtaaaacct ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgcgataat
     5101 tccgggccgc tttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     5161 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg
     5221 gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     5281 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcg
//
