LOCUS       sequence-377974-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EB3-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  AUTHORS   Stepanova T, Slemmer J, Hoogenraad CC, Lansbergen G, Dortland B, De 
            Zeeuw CI, Grosveld F, van Cappellen G, Akhmanova A, Galjart N
  TITLE     Visualization of microtubule growth in cultured neurons via the use 
            of EB3-GFP (end-binding protein 3-green fluorescent protein).
  JOURNAL   J Neurosci. 2003 Apr 1;23(7):2655-64.
  PUBMED    12684451
REFERENCE   2  (bases 1 to 5570)
  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: "J 
            Neurosci."; date: "2003-04-1"; volume: "23(7)"; pages: "2655-64"
COMMENT     Sequence Label: EB3-GFP
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1533..1542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1539..1541,1542..1544,1545..2258)
                     /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(1539..1541,1542..1544,1545..2255)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1584..1605)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1845..1864)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2192..2213
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2381..2502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2418..2437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2472..2491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2509..2964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2646..2667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2858..2877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2991..3095
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3097..3454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3097..3288
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3120..3140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3305..3440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3489..4283
                     /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             3489..4283
                     /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(3543..3562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4153..4172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4471..4490)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4515..4562
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4891..5479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5380..5399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      601 aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcaagctt cgaattctgc
      661 agtcgactgc caccatggcc gtcaatgtgt actccacatc tgtgaccagt gaaaatctga
      721 gtcgccatga tatgcttgca tgggtcaacg actccctgca cctcaactat accaagatag
      781 aacagctttg ttcaggggca gcctactgcc agttcatgga catgctcttc cccggctgtg
      841 tgcacttgag gaaagtgaag ttccaggcca aactagagca tgaatacatc cacaacttca
      901 aggtgctgca agcagctttc aagaagatgg gtgttgacaa aatcattcct gtagagaaat
      961 tagtgaaagg aaaattccaa gataattttg agtttattca gtggtttaag aaattctttg
     1021 acgcaaacta tgatggaaag gattacaacc ctctgctggc gcggcagggc caggacgtag
     1081 cgccacctcc taacccaggt gatcagatct tcaacaaatc caagaaactc attggcacag
     1141 cagttccaca gaggacgtcc cccacaggcc caaaaaacat gcagacctct ggccggctga
     1201 gcaatgtggc ccccccctgc attctccgga agaatcctcc atcagcccga aatggcggcc
     1261 atgagactga tgcccaaatt cttgaactca accaacagct ggtggacttg aagctgacag
     1321 tggatgggct ggagaaggaa cgtgacttct acttcagcaa acttcgtgac atcgagctca
     1381 tctgccagga gcatgaaagt gaaaacagcc ctgttatctc aggcatcatt ggcatcctct
     1441 atgccacaga ggaaggattc gcaccccctg aggacgatga gattgaagag catcaacaag
     1501 aagaccagga cgagtacctg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1561 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1621 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1681 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1741 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1801 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1861 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1921 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1981 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2041 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2101 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2161 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2221 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     2281 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2341 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2461 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2521 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2581 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2641 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2701 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2761 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2821 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2881 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2941 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3061 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3121 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3421 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3481 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3541 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3601 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3661 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3721 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3781 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3841 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3901 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3961 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4021 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4081 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4141 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4201 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4261 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4321 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4381 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4441 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4501 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4561 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4621 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4681 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4741 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4801 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4861 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4921 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4981 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5041 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5101 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5161 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5221 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5281 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5341 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5401 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5461 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5521 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
//
