LOCUS       Exported                5572 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of microtuble plus ends (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    EB3-mTurquoise2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5572)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv 160374
REFERENCE   2  (bases 1 to 5572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5572
                     /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      646..655
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      1510..1519
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1516..2235
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1561..1582)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1822..1841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2169..2190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2360..2481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2397..2416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2451..2470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2488..2943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2625..2646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2837..2856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2970..3074
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3076..3433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3099..3119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3284..3419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3346..3365
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3468..4262
                     /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(3522..3541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4132..4151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4450..4469)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4494..4541
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4870..5458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5359..5378
                     /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 cgactgccac catggccgtc
      661 aatgtgtact ccacatctgt gaccagtgaa aatctgagtc gccatgatat gcttgcatgg
      721 gtcaacgact ccctgcacct caactatacc aagatagaac agctttgttc aggggcagcc
      781 tactgccagt tcatggacat gctcttcccc ggctgtgtgc acttgaggaa agtgaagttc
      841 caggccaaac tagagcatga atacatccac aacttcaagg tgctgcaagc agctttcaag
      901 aagatgggtg ttgacaaaat cattcctgta gagaaattag tgaaaggaaa attccaagat
      961 aattttgagt ttattcagtg gtttaagaaa ttctttgacg caaactatga tggaaaggat
     1021 tacaaccctc tgctggcgcg gcagggccag gacgtagcgc cacctcctaa cccaggtgat
     1081 cagatcttca acaaatccaa gaaactcatt ggcacagcag ttccacagag gacgtccccc
     1141 acaggcccaa aaaacatgca gacctctggc cggctgagca atgtggcccc cccctgcatt
     1201 ctccggaaga atcctccatc agcccgaaat ggcggccatg agactgatgc ccaaattctt
     1261 gaactcaacc aacagctggt ggacttgaag ctgacagtgg atgggctgga gaaggaacgt
     1321 gacttctact tcagcaaact tcgtgacatc gagctcatct gccaggagca tgaaagtgaa
     1381 aacagccctg ttatctcagg catcattggc atcctctatg ccacagagga aggattcgca
     1441 ccccctgagg acgatgagat tgaagagcat caacaagaag accaggacga gtacctggat
     1501 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1561 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1621 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1681 gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt cgcccgctac
     1741 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1801 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1861 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1921 aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta tatcaccgcc
     1981 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     2041 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2101 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     2161 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2221 gagctgtaca agtaagaatt ggccgcgact ctagatcata atcagccata ccacatttgt
     2281 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2341 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2401 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2461 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2521 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2581 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2641 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2701 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2761 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2821 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2881 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3061 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3181 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3241 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3301 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3361 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3421 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3481 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3541 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3601 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3661 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3721 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3781 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3841 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3901 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3961 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4021 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4081 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4141 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4201 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4261 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4321 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4381 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4441 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4501 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4561 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4621 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4681 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5521 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
