LOCUS       sequence-256901-       11705 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor template to knock in HLA-E fused to B2M.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11705)
  TITLE     Lian Lab CRISPR plasmids
REFERENCE   2  (bases 1 to 11705)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLR04_AAVS1_Donor_HLAEB2M
FEATURES             Location/Qualifiers
     source          1..11705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    37..840
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    847..872
                     /label=SA
                     /note="splice acceptor site"
     CDS             896..949
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             959..1558
                     /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(959..978)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1455..1475
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1590..1607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1596..1820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1826..2205
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2207..2484
                     /label=chicken beta-actin promoter
     intron          2485..3501
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3425..3447
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3509..3528
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    5066..5654
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5119..5139)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5537..5548)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5686..6162
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5885..5904)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6272..6291)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6337..6392
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6391..6410)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6753..6769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6777..6793
                     /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)."
     promoter        complement(join(6801..6807,6808..6825,6826..6831))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6846..6867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6941..7777
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(7811..7830)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7812..7830)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7837..7854)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7837..7853)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7846..7868)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             7991..8293
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8172..8191
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8696..8715)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9310..9329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9690..10481,10482..10550))
                     /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     10313..10332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10674..11262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11163..11182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11416..11433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11550..11571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11586..11591,11592..11609,11610..11616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11624..11640
                     /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     11648..11664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11685..11703
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagctagtc ctgcaggttt aaacgaattc gccctttgct ttctctgacc agcattctct
       61 cccctgggcc tgtgccgctt tctgtctgca gcttgtggcc tgggtcacct ctacggctgg
      121 cccagatcct tccctgccgc ctccttcagg ttccgtcttc ctccactccc tcttcccctt
      181 gctctctgct gtgttgctgc ccaaggatgc tctttccgga gcacttcctt ctcggcgctg
      241 caccacgtga tgtcctctga gcggatcctc cccgtgtctg ggtcctctcc gggcatctct
      301 cctccctcac ccaaccccat gccgtcttca ctcgctgggt tcccttttcc ttctccttct
      361 ggggcctgtg ccatctctcg tttcttagga tggccttctc cgacggatgt ctcccttgcg
      421 tcccgcctcc ccttcttgta ggcctgcatc atcaccgttt ttctggacaa ccccaaagta
      481 ccccgtctcc ctggctttag ccacctctcc atcctcttgc tttctttgcc tggacacccc
      541 gttctcctgt ggattcgggt cacctctcac tcctttcatt tgggcagctc ccctaccccc
      601 cttacctctc tagtctgtgc tagctcttcc agccccctgt catggcatct tccaggggtc
      661 cgagagctca gctagtcttc ttcctccaac ccgggcccct atgtccactt caggacagca
      721 tgtttgctgc ctccagggat cctgtgtccc cgagctggga ccaccttata ttcccagggc
      781 cggttaatgt ggctctggtt ctgggtactt ttatctgtcc cctccacccc acagtggggc
      841 aagcttctga cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg
      901 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta ggctcgagat
      961 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     1021 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     1081 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     1141 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     1201 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     1261 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     1321 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     1381 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     1441 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     1501 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatc
     1561 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1621 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1681 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1741 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1801 ggatgcggtg ggctctatgg gtctcgacat tgattattga ctagttatta atagtaatca
     1861 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1921 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1981 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     2041 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     2101 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     2161 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
     2221 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     2281 attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccrggg
     2341 gggggggggg gggggggggg ggggggsggg gsgrggcgga gaggtgcggc ggcagccaat
     2401 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
     2461 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
     2521 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     2581 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
     2641 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
     2701 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
     2761 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
     2821 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
     2881 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
     2941 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     3001 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     3061 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     3121 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     3181 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
     3241 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
     3301 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     3361 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     3421 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     3481 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     3541 tttggcaaag aattcgtcga atggtagatg gaaccctcct tttactcctc tcggaggccc
     3601 tggcccttac ccagacctgg gcgggctccc actccttgaa gtatttccac acttccgtgt
     3661 cccggcccgg ccgcggggag ccccgcttca tctctgtggg ctacgtggac gacacccagt
     3721 tcgtgcgctt cgacaacgac gccgcgagtc cgaggatggt gccgcgggcg ccgtggatgg
     3781 agcaggaggg gtcagagtat tgggaccggg agacacggag cgccagggac accgcacaga
     3841 ttttccgagt gaatctgcgg acgctgcgcg gctactacaa tcagagcgag gccgggtctc
     3901 acaccctgca gtggatgcat ggctgcgagc tggggcccga cgggcgcttc ctccgcgggt
     3961 atgaacagtt cgcctacgac ggcaaggatt atctcaccct gaatgaggac ctgcgctcct
     4021 ggaccgcggt ggacacggcg gctcagatct ccgagcaaaa gtcaaatgat gcctctgagg
     4081 cggagcacca gagagcctac ctggaagaca catgcgtgga gtggctccac aaatacctgg
     4141 agaaggggaa ggagacgctg cttcacctgg agcccccaaa gacacacgtg actcaccacc
     4201 ccatctctga ccatgaggcc accctgaggt gctgggccct gggcttctac cctgcggaga
     4261 tcacactgac ctggcagcag gatggggagg gccataccca ggacacggag ctcgtggaga
     4321 ccaggcctgc aggggatgga accttccaga agtgggcagc tgtggtggtg ccttctggag
     4381 aggagcagag atacacgtgc catgtgcagc atgaggggct acccgagccc gtcaccctga
     4441 gatggaagcc ggcttcccag cccaccatcc ccatcgtggg catcattgct ggcctggttc
     4501 tccttggatc tgtggtctct ggagctgtgg ttgctgctgt gatatggagg aagaagagct
     4561 caggtggaaa aggagggagc tactctaagg ctgagtggag cgacagtgcc caggggtctg
     4621 agtctcacag cttgggagga ggtggaagcg gaggaggagg aagcggagga ggaggtagct
     4681 ctcgctccgt ggccttagct gtgctcgcgc tactctctct ttctggcctg gaggctatcc
     4741 agcgtactcc aaagattcag gtttactcac gtcatccagc agagaatgga aagtcaaatt
     4801 tcctgaattg ctatgtgtct gggtttcatc catccgacat tgaagttgac ttactgaaga
     4861 atggagagag aattgaaaaa gtggagcatt cagacttgtc tttcagcaag gactggtctt
     4921 tctatctctt gtactacact gaattcaccc ccactgaaaa agatgagtat gcctgccgtg
     4981 tgaaccatgt gactttgtca cagcccaaga tagttaagtg ggatcgagac atgtaacgcg
     5041 tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     5101 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5161 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5221 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5281 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5341 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5461 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     5521 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5581 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5641 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     5701 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     5761 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     5821 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     5881 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     5941 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     6001 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     6061 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     6121 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     6181 aggcgcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     6241 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     6301 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     6361 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     6421 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     6481 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     6541 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     6601 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     6661 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6721 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6781 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6841 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6901 ggaaacctgt cgtgccagcg gatcgacagt actaagcttt actagggaca ggattggtga
     6961 cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc
     7021 cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc
     7081 tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg
     7141 agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg
     7201 ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt
     7261 tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa
     7321 acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc
     7381 aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc
     7441 cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga
     7501 atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca
     7561 tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag
     7621 ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag
     7681 gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct
     7741 cctgcccctt ccctacaggg gttcctggct ctgctctaag ggcgaattcg cggccgctaa
     7801 attcaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     7861 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7921 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     7981 atacgtacgg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     8041 ggatgtacag agtgatatta ttgacacgcc ggggcgacgg atggtgatcc ccctggccag
     8101 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     8161 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     8221 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     8281 ctggggaata taaatgtcag gcatgagatt atcaaaaagg atcttcacct agatcctttt
     8341 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     8401 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     8461 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     8521 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     8581 aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag gatcgtttcg
     8641 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     8701 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     8761 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     8821 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     8881 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     8941 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     9001 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     9061 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     9121 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     9181 cggcgaggat ctcgtcgtga cccatgcatg gcgatgcctg cttgccgaat atcatggtgg
     9241 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     9301 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     9361 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     9421 ttcttgacga gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc
     9481 ttacgcatct gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc
     9541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagatta
     9601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     9781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    10021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    10081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    10141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    10201 actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc
    10261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    10321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    10381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    10441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    10501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    10561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgaccaaaat cccttaacgt
    10621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    10681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    10741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    10801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    10861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    10921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    10981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    11041 gaactgagat acctacagcg tgagctatga ggaagcgcca cgcttcccga agggagaaag
    11101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    11161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    11221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    11281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    11341 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    11401 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    11461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    11521 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    11581 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    11641 tttcacacag gaaacagcta tgaccatgat tacgccaagc tcagaattaa ccctcactaa
    11701 aggga
//
