LOCUS       Exported                5551 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-mScarlet-I
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  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 5551)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5551
                     /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..2211
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1649..1667)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1917..1936
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2339..2460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2376..2395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2430..2449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2467..2922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2816..2835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2949..3053
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3055..3412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3078..3098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3263..3398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3325..3344
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3447..4241
                     /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(3501..3520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4111..4130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4429..4448)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4473..4520
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4849..5437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5338..5357
                     /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 gaggcagtga tcaaggagtt catgcggttc
     1561 aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     1621 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1681 cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcatcaag
     1741 caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag
     1801 cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag
     1861 gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc
     1921 gtaatgcaga agaagacaat gggctgggaa gcgtccaccg agcggttgta ccccgaggac
     1981 ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg
     2041 gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac
     2101 gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac
     2161 gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattg
     2221 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2281 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2461 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2941 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3001 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3061 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3421 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3481 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3541 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3601 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3661 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3721 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3781 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3841 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3901 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3961 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4021 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4081 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4141 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4201 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4261 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4321 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4381 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4441 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4501 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4561 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4621 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4681 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4981 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5521 ttctgtggat aaccgtatta ccgccatgca t
//
