LOCUS       Exported                4919 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  mRuby2_PUFa in pmax-DEST expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    pAC1448_pmax-mRuby2_PUFa
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  AUTHORS   Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, 
            Taghbalout A, Wang H
  TITLE     Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation 
            and genomic labeling.
  JOURNAL   Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 
            Jan 15.
  PUBMED    26768771
REFERENCE   2  (bases 1 to 4919)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4919
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    960..984
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1002..1011
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1023..1040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1044..1076
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1080..1103
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1110..1820
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             1857..1877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(2949..2973)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3026..3067
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3093..3112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3122..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3147..3166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3401..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3481..3500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4069..4863)
                     /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(4180..4199)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4790..4809
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga
      961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg gtgcggggtt
     1021 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
     1081 atctgtacga cgatgacgat aaggatccga tggtgtctaa gggcgaagag ctgatcaagg
     1141 aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca
     1201 caggtgaagg agaaggcaat ccgtacatgg gaactcaaac catgaggatc aaagtcatcg
     1261 agggaggacc cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc
     1321 gtacttttat caagtacccg aaaggcattc ctgatttctt taaacagtcc tttcctgagg
     1381 gttttacttg ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcatgcagg
     1441 acaccagcct tgaggatggc tgtctcgttt accacgtcca agtcagaggg gtaaactttc
     1501 cctccaatgg tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga
     1561 tgtatccagc agatggtggt ctgaggggat acactcatat ggcactgaaa gttgatggtg
     1621 gtggccatct gtcttgctct ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca
     1681 tcaagatgcc cggtatccat gccgttgatc accgcctgga aaggttagag gaaagtgaca
     1741 atgaaatgtt cgtagtacaa cgcgaacacg cagttgccaa gttcgccggg cttggtggtg
     1801 ggatggacga gctgtacaag ggaggcggcg gtagcgggcc ggccgggatc ctccccccca
     1861 agaaaaagag gaaggtatct agaggccgca gccgcctttt ggaagatttt cgaaacaacc
     1921 ggtaccccaa tttacaactg cgggagattg ctggacatat aatggaattt tcccaagacc
     1981 agcatgggtc cagattcatt cagctgaaac tggagcgtgc cacaccagct gagcgccagc
     2041 ttgtcttcaa tgaaatcctc caggctgcct accaactcat ggtggatgtg tttggtaatt
     2101 acgtcattca gaagttcttt gaatttggca gtcttgaaca gaagctggct ttggcagaac
     2161 ggattcgagg ccacgtcctg tcattggcac tacagatgta tggcagccgt gttatcgaga
     2221 aagctcttga gtttattcct tcagaccagc agaatgagat ggttcgggaa ctagatggcc
     2281 atgtcttgaa gtgtgtgaaa gatcagaatg gcaatcacgt ggttcagaaa tgcattgaat
     2341 gtgtacagcc ccagtctttg caatttatca tcgatgcgtt taagggacag gtatttgcct
     2401 tatccacaca tccttatggc tgccgagtga ttcagagaat cctggagcac tgtctccctg
     2461 accagacact ccctatttta gaggagcttc accagcacac agagcagctg gtacaggatc
     2521 aatatggaaa ttatgtaatc caacatgtac tggagcacgg tcgtcctgag gataaaagca
     2581 aaattgtagc agaaatccga ggcaatgtac ttgtattgag tcagcacaaa tttgcaagca
     2641 atgttgtgga gaagtgtgtt actcacgcct cacgtacgga gcgcgctgtg ctcatcgatg
     2701 aggtgtgcac catgaacgac ggtccccaca gtgccttata caccatgatg aaggaccagt
     2761 atgccaacta cgtggtccag aagatgattg acgtggcgga gccaggccag cggaagatcg
     2821 tcatgcataa gatccggccc cacatcgcaa ctcttcgtaa gtacacctat ggcaagcaca
     2881 ttctggccaa gctggagaag tactacatga agaacggtgt tgacttaggg tgattaatta
     2941 agaattcgac ccagctttct tgtacaaagt ggttgatatc cagcacagtg gcggccgctc
     3001 gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3061 ttctacgcgt accggttagc aattgttttt tcgatgagtt tggacaaacc acaactagaa
     3121 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3181 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3241 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacttaag
     3301 agggggagac caaagggcga gacgttaagg cctcacgtga catgtgagca aaaggccagc
     3361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3421 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3481 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3541 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3841 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3901 gctcttgatc cggcaaacaa accacgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgccgtctc agaagaactc
     4081 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     4141 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     4201 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     4261 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     4321 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     4381 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     4441 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     4501 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     4561 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     4621 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4681 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4741 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4801 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4861 cataatatta ttgaagcatt tatcagggtt cgtctcgtcc cggtctcctc ccatgcatg
//
