LOCUS       sequence-330657-        8871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein Barcode (Pro-Code) for vector/cell tracking. Contains U6 
            trRNA cassette for sgRNA cloning. Enables phenotypic CRISPR screens 
            at a single cell resolution (using cytometry)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8871)
  AUTHORS   Wroblewska A, Dhainaut M, Ben-Zvi B, Rose SA, Park ES, Amir ED, 
            Bektesevic A, Baccarini A, Merad M, Rahman AH, Brown BD
  TITLE     Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens.
  JOURNAL   Cell. 2018 Nov 1;175(4):1141-1155.e16. doi: 
            10.1016/j.cell.2018.09.022. Epub 2018 Oct 18.
  PUBMED    30343902
REFERENCE   2  (bases 1 to 8871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2018.09.022"; journalName: "Cell"; date: "2018-11-1-
            1"; volume: "175"; issue: "4"; pages: "1141-1155.e16"
COMMENT     Sequence Label: pLKO_S.V5.VSVg_NGFR
FEATURES             Location/Qualifiers
     source          1..8871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     813..833
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      914..923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1016..1060
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1079..1120
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1139..1171
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     misc_feature    1934..2522
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1987..2007)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2405..2416)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2597..2830
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2902..3023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2939..2958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2993..3012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3218..3237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3219..3237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3247..3264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3247..3263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3256..3278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3405..3860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3492..3511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3702..3723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3886..3990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3991..4059,4060..4851)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4209..4228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5022..5610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5511..5530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5764..5781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5934..5939,5940..5957,5958..5964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5972..5988
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5977..5999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5996..6012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5996..6012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6031..6051
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6033..6051
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6079..6305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6306..6486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6533..6658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7151..7384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7569..7613
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7911..8159
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7911..7931
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8082..8101
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8178..8253
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    8301..8418
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8558..8871,1..865)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(8789..8871,1..856)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
       61 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
      121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
      181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
      241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
      301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
      361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
      421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
      481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
      541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
      601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
      661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
      721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
      781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
      841 ttttcttcca tttcaggtgt cgtgaggaat tctgcagtcg atcgacggta ccgcgggccc
      901 gggatccacg gtcgccacca tgggggcagg tgccaccggc cgcgccatgg acgggccgcg
      961 cctgctgctg ttgctgcttc tgggggtgtc ccttggaggt gccaaggagg catgcaagga
     1021 gaccgccgcc gccaagttcg agaggcagca catggacagc ggaggaggcg ggggaggggg
     1081 caaaccaatt ccaaacccac tgctgggact ggattccaca ggaggaggcg ggggagggta
     1141 tacagatatc gaaatgaata gactcggaaa gcccacaggc ctgtacacac acagcggtga
     1201 gtgctgcaaa gcctgcaacc tgggcgaggg tgtggcccag ccttgtggag ccaaccagac
     1261 cgtgtgtgag ccctgcctgg acagcgtgac gttctccgac gtggtgagcg cgaccgagcc
     1321 gtgcaagccg tgcaccgagt gcgtggggct ccagagcatg tcggcgccgt gcgtggaggc
     1381 cgacgacgcc gtgtgccgct gcgcctacgg ctactaccag gatgagacga ctgggcgctg
     1441 cgaggcgtgc cgcgtgtgcg aggcgggctc gggcctcgtg ttctcctgcc aggacaagca
     1501 gaacaccgtg tgcgaggagt gccccgacgg cacgtattcc gacgaggcca accacgtgga
     1561 cccgtgcctg ccctgcaccg tgtgcgagga caccgagcgc cagctccgcg agtgcacacg
     1621 ctgggccgac gccgagtgcg aggagatccc tggccgttgg attacacggt ccacaccccc
     1681 agagggctcg gacagcacag cccccagcac ccaggagcct gaggcacctc cagaacaaga
     1741 cctcatagcc agcacggtgg caggtgtggt gaccacagtg atgggcagct cccagcccgt
     1801 ggtgacccga ggcaccaccg acaacctcat ccctgtctat tgctccatcc tggctgctgt
     1861 ggttgtgggc cttgtggcct acatagcctt caagaggtgg aacaggggta tcctctagag
     1921 cggccgcgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1981 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2041 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2101 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2161 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2221 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2281 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2341 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2401 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2461 tcttccgcat cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2521 gcctgggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     2581 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga tctgcttttt
     2641 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2701 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     2761 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     2821 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2881 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     2941 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3001 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     3241 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3301 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3361 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     3421 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3481 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3541 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3601 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3661 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3721 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3781 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3841 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3901 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3961 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4021 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4081 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4141 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4201 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4261 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4321 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4381 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4441 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4501 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4561 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4621 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4681 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4741 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4801 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4861 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5161 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5401 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5641 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5701 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5761 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5821 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5881 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5941 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6001 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     6061 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     6121 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgg cgattggtgg
     6181 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     6241 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     6301 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6361 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6421 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6481 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     6541 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     6601 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     6661 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     6721 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     6781 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     6841 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     6901 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     6961 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     7021 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     7081 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     7141 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     7201 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     7261 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     7321 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     7381 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     7441 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     7501 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     7561 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     7621 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     7681 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     7741 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     7801 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     7861 ctcgacggta tcgatcacga gactagtatc gagcggccgc ccccttcacc gagggcctat
     7921 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     7981 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     8041 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     8101 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     8161 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     8221 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt aattctcgac ctcgagacaa
     8281 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     8341 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     8401 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggccgcgg
     8461 ctcgaggcga taagctttgc aaagatggat aaagttttaa acagagagga atctttgcag
     8521 ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     8581 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     8641 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     8701 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     8761 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
     8821 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc t
//
