LOCUS       Exported                5553 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Visualization of microtubules.
ACCESSION   .
VERSION     .
KEYWORDS    EMTB-mTurquoise2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5553)
  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 5553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5553
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      271..280
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             280..309
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1129..1138
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1135..1854
                     /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(1180..1201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1441..1460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1788..1809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1977..2098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2014..2033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2068..2087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2105..2560)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2242..2263)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2454..2473
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2587..2691
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2693..3050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2716..2736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2901..3036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2963..2982
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3085..3879
                     /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(3139..3158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3749..3768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4067..4086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4111..4158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4487..5075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4976..4995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5250..5553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc gccaccatgg agcagaagct catctcagaa
      301 gaagacctcg gcgcgccagt gcgaagcgaa acagcacccg acagctacaa agtgcaagat
      361 aagaaaaatg cctccagccg ccctgcctct gcaatttcag gacaaaataa caaccactca
      421 ggaaataaac cagaccctcc gcctgtgtta cgtgttgatg accggcagcg gctggcccgg
      481 gagcgacgtg aggaacggga gaaacagcta gctgcaagag aaatagtgtg gttagaaaga
      541 gaagagcgag ccaggcagca ctacgagaag cacctggaag agcggaagaa gaggttggag
      601 gagcagaggc agaaggagga gcggaggagg gctgctgtgg aggagaagcg gaggcagaga
      661 cttgaggagg acaaagaacg ccacgaagct gttgtacggc gcacaatgga aaggagccag
      721 aagccaaaac agaagcataa ccgttggtcg tggggaggct ctctccatgg gagccctagc
      781 atccacagtg cagatccaga caggcggtca gtttccacca tgaatctttc gaaatatgtt
      841 gatcccgtca ttagcaagcg gctctcctct tcatctgcaa ctttactaaa ttctccagat
      901 agagctcgcc gcctgcagct cagcccatgg gagagcagcg ttgttaacag actcctgacg
      961 cccacacatt cgttcctggc cagaagtaaa agcacagctg ccttgtctgg agaagcagca
     1021 tcttgcagcc ccatcatcat gccctacaaa gctgcacact ctagaaattc gatggatcga
     1081 ccaaaactct ttgtaacacc acctgagggc tcttcggatc caccggtcgc caccatggtg
     1141 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1201 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1261 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1321 accaccctgt cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1381 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1441 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1501 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1561 gagtacaact actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1621 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1681 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1741 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1801 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     1861 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1921 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1981 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2041 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     2101 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2161 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2221 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2281 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2341 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2401 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2461 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2521 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2581 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2641 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2701 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3061 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3241 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3601 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4021 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4141 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4201 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4261 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4321 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5161 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5521 tacatctacg tattagtcat cgctattacc atg
//
