LOCUS       Exported                6280 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses EB3-tdTomato fusion in mammalian cells for labeling the 
            growing tips of MTs.
ACCESSION   .
VERSION     .
KEYWORDS    EB3-tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6280)
  AUTHORS   Merriam EB, Millette M, Lumbard DC, Saengsawang W, Fothergill T, Hu 
            X, Ferhat L, Dent EW
  TITLE     Synaptic regulation of microtubule dynamics in dendritic spines by 
            calcium, F-actin, and drebrin.
  JOURNAL   J Neurosci. 2013 Oct 16;33(42):16471-82. doi: 
            10.1523/JNEUROSCI.0661-13.2013.
  PUBMED    24133252
REFERENCE   2  (bases 1 to 6280)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6280
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      653..662
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      1517..1526
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1523..2953
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    3075..3196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3112..3131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3166..3185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3203..3658)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3340..3361)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3552..3571
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3685..3789
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3791..4148
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3814..3834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3999..4134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4061..4080
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4183..4977
                     /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(4237..4256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4847..4866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5165..5184)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5209..5256
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5585..6173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6074..6093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga ctgccaccat
      661 ggccgtcaat gtgtactcca catctgtgac cagtgaaaat ctgagtcgcc atgatatgct
      721 tgcatgggtc aacgactccc tgcacctcaa ctataccaag atagaacagc tttgttcagg
      781 ggcagcctac tgccagttca tggacatgct cttccccggc tgtgtgcact tgaggaaagt
      841 gaagttccag gccaaactag agcatgaata catccacaac ttcaaggtgc tgcaagcagc
      901 tttcaagaag atgggtgttg acaaaatcat tcctgtagag aaattagtga aaggaaaatt
      961 ccaagataat tttgagttta ttcagtggtt taagaaattc tttgacgcaa actatgatgg
     1021 aaaggattac aaccctctgc tggcgcggca gggccaggac gtagcgccac ctcctaaccc
     1081 aggtgatcag atcttcaaca aatccaagaa actcattggc acagcagttc cacagaggac
     1141 gtcccccaca ggcccaaaaa acatgcagac ctctggccgg ctgagcaatg tggccccccc
     1201 ctgcattctc cggaagaatc ctccatcagc ccgaaatggc ggccatgaga ctgatgccca
     1261 aattcttgaa ctcaaccaac agctggtgga cttgaagctg acagtggatg ggctggagaa
     1321 ggaacgtgac ttctacttca gcaaacttcg tgacatcgag ctcatctgcc aggagcatga
     1381 aagtgaaaac agccctgtta tctcaggcat cattggcatc ctctatgcca cagaggaagg
     1441 attcgcaccc cctgaggacg atgagattga agagcatcaa caagaagacc aggacgagta
     1501 cctggatcca ccggtcgcca ccatggtgag caagggcgag gaggtcatca aagagttcat
     1561 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     1621 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     1681 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     1741 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     1801 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     1861 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     1921 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     1981 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     2041 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     2101 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2161 acagtacgag cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac
     2221 cggcagcggc agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga
     2281 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga
     2341 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2401 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
     2461 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
     2521 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
     2581 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
     2641 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     2701 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
     2761 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
     2821 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
     2881 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga
     2941 gctgtacaag taggcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3001 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3061 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3181 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3241 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3301 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3361 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3421 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3481 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3541 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3601 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3781 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4081 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4141 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5041 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5101 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5221 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5281 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5341 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5401 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5461 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5701 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5761 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5821 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5881 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5941 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6001 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6061 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6121 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6181 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6241 ctgcgttatc ccctgattct gtggataacc gtattaccgc
//
