                         SEQUENCE LISTING

<110>  The Broad Institute
       Blainey, Paul
       Borrajo, Jacob
       Najia, Mohamad
       Kulshreshtha, Ankur
       Dixit, Atray
 
<120>  Constructs for Continuous Monitoring of Live Cells

<130>  BROD-0820WP

<150>  US 62/397,867
<151>  2016-09-21

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  1590
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  MLV gag

<400>  1
atgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag       60

cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct      120

gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc      180

atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc      240

ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt      300

gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct      360

cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc      420

aaacctaaac ctcaagttct ttctgacagt ggggggccgc tcatcgacct acttacagaa      480

gaccccccgc cttataggga cccaagacca cccccttccg acagggacgg aaatggtgga      540

gaagcgaccc ctgcgggaga ggcaccggac ccctccccaa tggcatctcg cctacgtggg      600

agacgggagc cccctgtggc cgactccact acctcgcagg cattccccct ccgcgcagga      660

ggaaacggac agcttcaata ctggccgttc tcctcttctg acctttacaa ctggaaaaat      720

aataaccctt ctttttctga agatccaggt aaactgacag ctctgatcga gtctgttctc      780

atcacccatc agcccacctg ggacgactgt cagcagctgt tggggactct gctgaccgga      840

gaagaaaaac aacgggtgct cttagaggct agaaaggcgg tgcggggcga tgatgggcgc      900

cccactcaac tgcccaatga agtcgatgcc gcttttcccc tcgagcgccc agactgggat      960

tacaccaccc aggcaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt     1020

ctccaaaacg cgggcagaag ccccaccaat ttggccaagg taaaaggaat aacacaaggg     1080

cccaatgagt ctccctcggc cttcctagag agacttaagg aagcctatcg caggtacact     1140

ccttatgacc ctgaggaccc agggcaagaa actaatgtgt ctatgtcttt catttggcag     1200

tctgccccag acattgggag aaagttagag aggttagaag atttaaaaaa caagacgctt     1260

ggagatttgg ttagagaggc agaaaagatc tttaataaac gagaaacccc ggaagaaaga     1320

gaggaacgta tcaggagaga aacagaggaa aaagaagaac gccgtaggac agaggatgag     1380

cagaaagaga aagaaagaga tcgtaggaga catagagaga tgagcaagct attggccact     1440

gtcgttagtg gacagaaaca ggatagacag ggaggagaac gaaggaggtc ccaactcgat     1500

cgcgaccagt gtgcctactg caaagaaaag gggcactggg ctaaagattg tcccaagaaa     1560

ccacgaggac ctcggggacc aagaccgcag                                      1590


<210>  2
<211>  1935
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PABPC4 - RNA binding protein fragment

<400>  2
atgaacgctg cggccagcag ctaccccatg gcctccctgt acgtgggcga cctgcattcg       60

gacgtcaccg aggccatgct gtacgaaaag ttcagccccg cggggcctgt gctgtccatc      120

cgggtctgcc gcgatatgat cacccgccgc tccctgggct atgcctacgt caacttccag      180

cagccggccg acgctgagcg ggctttggac accatgaact ttgatgtgat taagggaaag      240

ccaatccgca tcatgtggtc tcagagggat ccctctttga gaaaatctgg tgtgggaaac      300

gtcttcatca agaacctgga caaatctata gataacaagg cactttatga tactttttct      360

gcttttggaa acatactgtc ctgcaaggtg gtgtgtgatg agaacggctc taagggttat      420

gcctttgtcc acttcgagac ccaagaggct gccgacaagg ccatcgagaa gatgaatggc      480

atgctcctca atgaccgcaa agtatttgtg ggcagattca agtctcgcaa agagcgggaa      540

gctgagcttg gagccaaagc caaggaattc accaatgttt atatcaaaaa ctttggggaa      600

gaggtggatg atgagagtct gaaagagcta ttcagtcagt ttggtaagac cctaagtgtc      660

aaggtgatga gagatcccaa tgggaaatcc aaaggctttg gctttgtgag ttacgaaaaa      720

cacgaggatg ccaataaggc tgtggaagag atgaatggaa aagaaataag tggtaaaatc      780

atatttgtag gccgtgcaca aaagaaagta gaacggcagg cagagttaaa acggaaattt      840

gaacagttga aacaggagag aattagtcga tatcaggggg tgaatctcta cattaagaac      900

ttggatgaca ctattgatga tgagaaatta aggaaagaat tttctccttt tggatcaatt      960

accagtgcta aggtaatgct ggaggatgga agaagcaaag ggtttggctt cgtctgcttc     1020

tcatctcctg aagaagcaac caaagcagtc actgagatga atggacgcat tgtgggctcc     1080

aagccactat atgttgccct ggcccagagg aaggaagaga gaaaggctca cctgaccaac     1140

cagtatatgc aacgagtggc tggaatgaga gcacttcctg ccaatgccat cttaaatcag     1200

ttccagcctg cagcgggtgg ctactttgtg ccagcagtcc cacaggctca gggaaggcct     1260

ccatattata cacctaacca gttagcacag atgaggccta atccacgctg gcagcaaggt     1320

gggagacctc aaggcttcca aggaatgcca agtgctatac gccagtctgg gcctcgtcca     1380

actcttcgcc atctggctcc aactgggtct gagtgcccgg accgcttggc tatggacttt     1440

ggtggggctg gtgccgccca gcaagggctg actgacagct gccagtctgg aggcgttccc     1500

acagctgtgc agaacttagc gccacgcgct gctgttgctg ctgctgctcc ccgggctgtt     1560

gccccctaca aatacgcctc cagtgtccgc agccctcatc ctgccataca gcctctgcag     1620

gcaccccagc ctgcggtcca tgtgcagggg caggagccac tgactgcctc catgctggct     1680

gcagcacccc cccaggaaca gaagcagatg ctgggagaac gcttgttccc actcatccaa     1740

acaatgcatt caaatctggc tgggaagatc acgggaatgc tgctggagat agacaactct     1800

gagctgctgc acatgttaga gtcccccgag tctctccgct ccaaggtgga tgaagctgta     1860

gcagttctac aggctcatca tgccaagaaa gaagctgccc agaaggtggg cgctgttgct     1920

gctgctacct cttag                                                      1935


<210>  3
<211>  387
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  MS2 (N55K)  - RNA binding protein fragment

<400>  3
gcttcaaact ttactcagtt cgtgctcgtg gacaatggtg ggacagggga tgtgacagtg       60

gctccttcta atttcgctaa tggggtggca gagtggatca gctccaactc acggagccag      120

gcctacaagg tgacatgcag cgtcaggcag tctagtgccc agaagagaaa gtataccatc      180

aaggtggagg tccccaaagt ggctacccag acagtgggcg gagtcgaact gcctgtcgcc      240

gcttggaggt cctacctgaa catggagctc actatcccaa ttttcgctac caattctgac      300

tgtgaactca tcgtgaaggc aatgcagggg ctcctcaaag acggtaatcc tatcccttcc      360

gccatcgccg ctaactcagg tatctac                                          387


<210>  4
<211>  4137
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dCas9 -  - RNA binding protein fragment

<400>  4
atgagcccca agaagaagag aaaggtggag gccagcgaca agaagtacag catcggcctg       60

gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc      120

aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga      180

gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga      240

agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag      300

atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag      360

gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac      420

gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc      480

gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg      540

atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg      600

cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag      660

gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg      720

cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc      780

cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac      840

acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg      900

tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac      960

accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac     1020

caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag     1080

attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag     1140

gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg     1200

ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc     1260

atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt     1320

tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc     1380

tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc     1440

gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag     1500

agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc     1560

aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac     1620

gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg     1680

gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc     1740

aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc     1800

tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat     1860

gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga     1920

gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag     1980

cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc     2040

atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc     2100

aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag     2160

aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc     2220

agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa     2280

gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc     2340

acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa     2400

gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag     2460

aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac     2520

atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac     2580

gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac     2640

gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc     2700

aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc     2760

gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag     2820

cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg     2880

atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat     2940

ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg     3000

aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg     3060

tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc     3120

ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag     3180

attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc     3240

ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg     3300

ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct     3360

atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag     3420

aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg     3480

gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg     3540

gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa     3600

gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc     3660

cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc     3720

tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc     3780

gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc     3840

atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa     3900

gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc     3960

atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc     4020

accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac     4080

cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgac        4137


<210>  5
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  p2A protein domain

<400>  5
ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct       60

ggaccg                                                                  66


<210>  6
<211>  711
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  YFP  protein domain

<400>  6
agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac       60

gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag      120

ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg      180

accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac      240

gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag      300

gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac      360

cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg      420

gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc      480

aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac      540

taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg      600

agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg      660

gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa g               711


<210>  7
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GS6 linker

<400>  7
ggatcaggat caggatcagg a                                                 21


<210>  8
<211>  204
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  bGH pA terminator

<400>  8
cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga       60

ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt      120

gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg      180

attgggaaga caatagcagg catg                                             204


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  spacer

<400>  9
ctgcaggtcg actctagaaa                                                   20


<210>  10
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  MS2 hairpin

<400>  10
acatgaggat cacccatgt                                                    19


<210>  11
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Smartseq2 handle

<400>  11
aagcagtggt atcaacgcag agtac                                             25


<210>  12
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  polyA capture

<400>  12
ttgttgttgt tgttgttgtt tttt                                              24


