LOCUS       sequence-422312-       10947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2 hairpin-containing expression vector and sgRNA entry plasmid for
            the directed export of barcode transcripts in Gag-MCP virus-like 
            particles.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10947)
  AUTHORS   Najia MA, Borrajo J, Le A, Tsai F, Huang JY, Griffith LG, Daley GQ, 
            Blainey PC
  TITLE     Live-cell transcriptomics with engineered virus-like particles
  JOURNAL   bioRxiv 2024.10.01.616098
REFERENCE   2  (bases 1 to 10947)
  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"; journalName: "bioRxiv 
            2024.10.01.616098"
COMMENT     Sequence Label: CROPseq-Puro-12xMS2
FEATURES             Location/Qualifiers
     source          1..10947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /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"
     misc_feature    2003..2120
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2259..3437
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2490..3428
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3385..3405
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3450..4049
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3450..3469)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3946..3966
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_RNA        4106..4124
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4176..4194
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4215..4233
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4285..4303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4324..4342
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4394..4412
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4464..4482
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4503..4521
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4573..4591
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4612..4630
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4682..4700
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4721..4739
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    4788..5376
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4841..4861)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5259..5270)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5532..5780
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5532..5552
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5703..5722
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             7800..7980
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    8058..8179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8095..8114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8149..8168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8219..8354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8374..8393)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8375..8393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8403..8420)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8403..8419)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8412..8434)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8561..9016
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8648..8667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8858..8879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9042..9146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9147..9215,9216..10007)
                     /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(9365..9384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10178..10766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10667..10686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10920..10937
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgac
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga atcttgagac
     1981 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2041 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2101 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
     2161 ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca
     2221 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2281 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2341 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2401 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2461 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2521 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     2581 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     2641 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     2701 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     2761 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     2821 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     2881 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     2941 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     3001 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     3061 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     3121 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     3181 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     3241 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3301 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3361 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3421 catttcaggt gtcgtgacgt acggccacca tgaccgagta caagcccacg gtgcgcctcg
     3481 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3541 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3601 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3661 ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3721 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3781 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     3841 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     3901 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     3961 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4021 ggtgcatgac ccgcaagccc ggtgcctgat ggacgagctg tacaagtaaa gcggccggat
     4081 cctaaggtac ctaattgcct agaaaacatg aggatcaccc atgtctgcag tattcccggg
     4141 ttcattagat cctaaggtac ctaattgcct agaaaacatg aggatcaccc atgtctgcag
     4201 gtcgactcta gaaaacatga ggatcaccca tgtctgcagt attcccgggt tcattagatc
     4261 ctaaggtacc taattgccta gaaaacatga ggatcaccca tgtctgcagg tcgactccag
     4321 aaaacatgag gatcacccat gtctgcagta ttcccgggtt cattagatcc taaggtacct
     4381 aattgcctag aaaacatgag gatcacccat gtctgcagta ttcccgggtt cattagatcc
     4441 taaggtacct aattgcctag aaaacatgag gatcacccat gtctgcaggt cgactctaga
     4501 aaacatgagg atcacccatg tctgcagtat tcccgggttc attagatcct aaggtaccta
     4561 attgcctaga aaacatgagg atcacccatg tctgcaggtc gactctagaa aacatgagga
     4621 tcacccatgt ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa
     4681 aacatgagga tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtc
     4741 tgcagtattc ccgggttcat tagatctgcg cgcgatcgat agatcctaat caacctctgg
     4801 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4861 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4921 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4981 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5041 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5101 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5161 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5221 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5281 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5341 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta
     5401 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5461 tcactcccaa cgaagacaag atttcaccat tatcgtttca gacccacctc ccaaccccga
     5521 ggggacccag agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     5581 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     5641 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     5701 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     5761 tgtggaaagg acgaaacacc ggagacggtt gtaaatgagc acacaaaata cacatgctaa
     5821 aatattatat tctatgacct ttataaaatc aaccaaaatc ttctttttaa taactttagt
     5881 atcaataatt agaattttta tgttcctttt tgcaaacttt taataaaaat gagcaaaata
     5941 aaaaaacgct agttttagta actcgcgttg ttttcttcac ctttaataat agctactcca
     6001 ccacttgttc ctaagcggtc agctcctgct tcaatcattt tttgagcatc ttcaaatgtt
     6061 ctaactccac cagctgcttt aactaaagca ttgtctttaa caactgactt cattagttta
     6121 acatcttcaa atgttgcacc tgattttgaa aatcctgttg atgttttaac aaattctaat
     6181 ccagcttcaa cagctatttc acaagctttc atgatttctt cttttgttaa taaacaattt
     6241 tccataatac atttaacaac atgtgatcca gctgcttttt ttacagcttt catgtcttct
     6301 aaaactaatt cataattttt gtcttttaat gcaccaatat ttaataccat atcaatttct
     6361 gttgcaccat ctttaattgc ttcagaaact tcgaatgctt ttgtagctgt tgtgcatgca
     6421 cctagaggaa aacctacaac atttgttatt cctacatttg tgccttttaa taattcttta
     6481 caatagcttg ttcaatatga attaacacaa actgttgcaa aatcaaattc aattgcttca
     6541 tcacataatt gtttaatttc agctttcgta gcatcttgtt ttaataatgt gtgatctata
     6601 tatttgttta gtttcatttt ttctcctata tattcatttt taattttaat tctttaataa
     6661 tttcgtctac tttaacttta gcgttttgaa cagattcacc aacacctata aaataaattt
     6721 ttagtttagg ttcagttcca cttgggcgaa cagcaaatca tgacttatct tctaaataaa
     6781 attttagtaa gtcttgtcct ggcatattat acattccatc gatgtagtct tcaacattaa
     6841 caactttaag tccagcaatt tgagttaagg gtgttgctct caatgatttc attaatggtt
     6901 caatttttaa tttcttttct tctggtttaa aattcaagtt taaagtgaaa gtgtaatatg
     6961 cacccatttc tttaaataaa tcttctaaat agtctactaa tgttttattt tgttttttat
     7021 aaaatcaagc agcctctgct attaatatag aagcttgtat tccatcttta tctctagctg
     7081 agtcatcaat tacatatcca taactttctt cataagcaaa aacaaaattt aatccgttat
     7141 cttcttcttt agcaatttct ctacccattc atttaaatcc agttaaagtt tttacaatat
     7201 taactccata tttttcatga gcgattctat cacccaaatc acttgttaca aaacttgaat
     7261 atagagccgg attttttgga atgctattta agcgttttag atttgataat tttcaatcaa
     7321 ttaaaattgg tcctgtttga tttccatcta atcttacaaa atgaccatca tgttttattg
     7381 ccattccaaa tctgtcagca tctgggtcat tcataataat aatatctgca tcatgtttaa
     7441 taccatattc aagcggtatt tttcatgcag gatcaaattc tggatttgga tttacaacat
     7501 ttttaaatgt ttcatcttca aatgcatgct cttcaacctc aataacgtta tatcctgatt
     7561 cacgtaatat ttttggggta aatttagttc ctgttccatt aactgcgcta aaaataattt
     7621 ttaaatcttt tttagcttct tgctcttttt tgtacgtctc tgtttcagag ctatgctgga
     7681 aacagcatag caagttgaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     7741 tcggtgcttt tttaagcttg gcgtaactag atcttgagac actgcttttt gcttgtactg
     7801 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7861 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7921 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7981 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     8041 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     8101 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     8161 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     8221 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     8281 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     8341 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     8401 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     8461 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     8521 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     8581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     8641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     8701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     9001 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     9061 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     9121 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     9181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     9241 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     9301 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     9361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     9421 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     9481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     9541 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     9601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     9661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     9721 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     9781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     9841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     9901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     9961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
    10021 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
    10081 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
    10141 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
    10201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
    10261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
    10321 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
    10381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    10441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
    10501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
    10561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
    10621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
    10681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
    10741 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
    10801 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
    10861 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
    10921 gcgagtcagt gagcgaggaa gcggaag
//
