LOCUS       Exported                4772 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of the nucleus (can be used for colocalization
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    3xnls-mScarlet-I
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4772)
  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 4772)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4772
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     239..259
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(284..388)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      415..870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(502..521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     712..733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    877..998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(888..907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     942..961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1124..1819)
                     /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(1399..1418)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1668..1686
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1816..1825
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(1838..1858)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1862..1882)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1886..1906)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1990..2193)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2019..2039)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2194..2497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2672..3260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2752..2771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3589..3636
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3661..3680
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3868..4662)
                     /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(3979..3998)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4589..4608
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4697..282)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4711..74
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4765..12)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
       61 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
      121 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
      181 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      241 cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
      301 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
      361 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
      421 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      481 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      541 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      601 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      661 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      721 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      781 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      841 attttaacaa aatattaacg cttacaattt acgccttaag atacattgat gagtttggac
      901 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      961 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1021 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1081 aatgtggtat ggctgattat gatctagagt cgcggccgct ttacttgtac agctcgtcca
     1141 tgccgccggt ggagtggcgg ccctcggagc gttcgtactg ttccaccacg gtgtagtcct
     1201 cgttgtggga ggtgatgtcc aacttgcggt cgacgttgta ggcgccgggc atctgcacgg
     1261 gcttcttggc cttgtaggtg gtcttgaagt ccgccaggta gcggccgccg tccttcaggc
     1321 gcagggccat cttaatgtcg cccttcagca cgccgtcctc ggggtacaac cgctcggtgg
     1381 acgcttccca gcccattgtc ttcttctgca ttacggggcc gtcaggaggg aagttggtgc
     1441 cgcggagctt caccttgtag atcagggtgc cgtcctccag ggaggtgtcc tgggtcacgg
     1501 tcacggcgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     1561 actgcttata gtagtcgggg atgtcggcgg ggtgcttgat gaaggccctg gagccgtaca
     1621 tgaactgagg ggacaggatg tcccaggaga agggcagggg gccacccttg gtcaccttca
     1681 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     1741 cgtggccgtt catggagccc tccatgtgca ccttgaaccg catgaactcc ttgatcactg
     1801 cctcgccctt gctcaccatg gtggcgaccg gtgatcccac tttccgtttc tttttgggat
     1861 cgaccttacg ctttttcttc gggtctacct ttctcttctt ttttggatct gatcccatgg
     1921 tgggaattcg aagcttgagc tcgagatctg agtccggtag cgctagcgga tctgacggtt
     1981 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2041 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2101 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2161 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2221 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2281 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2341 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2401 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2461 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2521 aactaatgac cccgtaattg attactatta ataactaatg catggcggta atacggttat
     2581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     3421 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     3481 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     3541 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     3601 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     3661 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     3721 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     3781 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     3841 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     3901 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     3961 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     4021 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     4081 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     4141 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     4201 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     4261 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     4321 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     4381 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     4441 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     4501 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     4561 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     4621 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     4681 tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     4741 ggaatagctc agaggccgag gcggcctcgg cc
//
