LOCUS       sequence-308780-       11501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock CLTX-CAR into AAVS1 safe harbor locus for constitutive 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11501)
  AUTHORS   Chang Y, Syahirah R, Wang X, Jin G, Torregrosa-Allen S, Elzey BD, 
            Hummel SN, Wang T, Li C, Lian X, Deng Q, Broxmeyer HE, Bao X
  TITLE     Engineering chimeric antigen receptor neutrophils from human 
            pluripotent stem cells for targeted cancer immunotherapy.
  JOURNAL   Cell Rep. 2022 Jul 19;40(3):111128. doi: 
            10.1016/j.celrep.2022.111128.
  PUBMED    35858579
REFERENCE   2  (bases 1 to 11501)
  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: "Cell Rep.";
            date: "2022-07-19"; volume: "40(3):111128. doi"; pages: " 
            10.1016/j.celrep.2022.111128"
COMMENT     Sequence Label: AAVS1-Puro CAG CLTX-CAR
FEATURES             Location/Qualifiers
     source          1..11501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..836
                     /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    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /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             955..1554
                     /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(955..974)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1451..1471
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1586..1603)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1822..2201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2203..2480
                     /label=chicken beta-actin promoter
     intron          2481..3497
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3421..3443
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3505..3524
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    4863..5451
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4916..4936)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5334..5345)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5483..5959
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5682..5701)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6069..6088)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6134..6189
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6188..6207)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6550..6566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6574..6590
                     /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(6598..6604,6605..6622,6623..6628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6643..6664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6738..7574
                     /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(7608..7627)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7609..7627)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7634..7651)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7634..7650)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7643..7665)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             7788..8090
                     /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     7969..7988
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8493..8512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9107..9126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9487..10278,10279..10347))
                     /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     10110..10129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10471..11059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10960..10979
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11213..11230
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11347..11368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11383..11388,11389..11406,11407..11413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11421..11437
                     /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     11445..11461
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11482..11500
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc
       61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca
      121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc
      181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc
      241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc
      301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg
      361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc
      421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc
      481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc
      541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta
      601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag
      661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt
      721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt
      781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc
      841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga
      901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagatgacc
      961 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     1021 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     1081 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1141 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1201 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1261 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1321 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     1381 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1441 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1501 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatctaga
     1561 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1621 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1681 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1741 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1801 gcggtgggct ctatgggtct cgacattgat tattgactag ttattaatag taatcaatta
     1861 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1921 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1981 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2041 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2101 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2161 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
     2221 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
     2281 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccrggcgggg
     2341 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     2401 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     2461 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     2521 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     2581 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     2641 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     2701 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
     2761 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     2821 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     2881 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     2941 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     3001 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     3061 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     3121 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     3181 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     3241 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     3301 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     3361 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     3421 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     3481 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     3541 gcaaagaatt cgtcgacgcc accatgcttc tcctggtgac aagccttctg ctctgtgagt
     3601 taccacaccc agcattcctc ctgatcccaa tgtgcatgcc ttgcttcacc accgaccacc
     3661 agatggccag aaagtgcgat gactgctgcg gcggcaaggg cagaggcaag tgctacggcc
     3721 cccagtgtct gtgcagagag tccaaatatg gtcccccatg cccaccatgc ccagcacctg
     3781 agttcctggg gggaccatca gtcttcctgt tccccccaaa acccaaggac actctcatga
     3841 tctcccggac ccctgaggtc acgtgcgtgg tggtggacgt gagccaggaa gaccccgagg
     3901 tccagttcaa ctggtacgtg gatggcgtgg aggtgcataa tgccaagaca aagccgcggg
     3961 aggagcagtt caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact
     4021 ggctgaacgg caaggagtac aagtgcaagg tctccaacaa aggcctcccg tcctccatcg
     4081 agaaaaccat ctccaaagcc aaagggcagc cccgagagcc acaggtgtac accctgcccc
     4141 catcccagga ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct
     4201 accccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga
     4261 ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcagg ctaaccgtgg
     4321 acaagagcag gtggcaggag gggaatgtct tctcatgctc cgtgatgcat gaggctctgc
     4381 acaaccacta cacacagaag agcctctccc tgtccctagg taaaatggcc ctgattgtgc
     4441 tggggggcgt cgccggcctc ctgcttttca ttgggctagg catcttcttc agagtgaagt
     4501 tcagcaggag cgcagacgcc cccgcgtacc agcagggcca gaaccagcag ctctataacg
     4561 agctcaatct aggacgaaga gaggagtacg atgttttgga caagagacgt ggccgggacc
     4621 ctgagatggg gggaaagccg agaaggaaga accctcagga aggcctgtac aatgaactgc
     4681 agaaagataa gatggcggag gcctacagtg agattgggat gaaaggcgag cgccggaggg
     4741 gcaaggggca cgatggcctt taccagggtc tcagtacagc caccaaggac acctacgacg
     4801 cccttcacat gcaggccctg ccccctcgct gaacgcgtga attcgatatc aagcttatcg
     4861 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4921 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4981 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5041 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5101 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5161 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5221 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5281 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5341 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5401 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     5461 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     5521 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     5581 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     5641 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     5701 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     5761 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     5821 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     5881 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     5941 tgctcccttc cctgtccttc tgattttgta ggtaaccagg cgcgtgaatt cactcctcag
     6001 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     6061 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     6121 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     6181 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     6241 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     6301 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     6361 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     6421 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     6481 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     6541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     6721 cgacagtact aagctttact agggacagga ttggtgacag aaaagcccca tccttaggcc
     6781 tcctccttcc tagtctcctg atattgggtc taacccccac ctcctgttag gcagattcct
     6841 tatctggtga cacaccccca tttcctggag ccatctctct ccttgccaga acctctaagg
     6901 tttgcttacg atggagccag agaggatcct gggagggaga gcttggcagg gggtgggagg
     6961 gaaggggggg atgcgtgacc tgcccggttc tcagtggcca ccctgcgcta ccctctccca
     7021 gaacctgagc tgctctgacg cggctgtctg gtgcgtttca ctgatcctgg tgctgcagct
     7081 tccttacact tcccaagagg agaagcagtt tggaaaaaca aaatcagaat aagttggtcc
     7141 tgagttctaa ctttggctct tcacctttct agtccccaat ttatattgtt cctccgtgcg
     7201 tcagttttac ctgtgagata aggccagtag ccagccccgt cctggcaggg ctgtggtgag
     7261 gaggggggtg tccgtgtgga aaactccctt tgtgagaatg gtgcgtccta ggtgttcacc
     7321 aggtcgtggc cgcctctact ccctttctct ttctccatcc ttctttcctt aaagagtccc
     7381 cagtgctatc tgggacatat tcctccgccc agagcagggt cccgcttccc taaggccctg
     7441 ctctgggctt ctgggtttga gtccttggca agcccaggag aggcgctcag gcttccctgt
     7501 cccccttcct cgtccaccat ctcatgcccc tggctctcct gccccttccc tacaggggtt
     7561 cctggctctg ctctaagggc gaattcgcgg ccgctaaatt caattcgccc tatagtgagt
     7621 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     7681 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     7741 cccgcaccga tcgcccttcc caacagttgc gcagcctata cgtacggcag tttaaggttt
     7801 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     7861 acacgccggg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     7921 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     7981 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     8041 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggca
     8101 tgagattatc aaaaaggatc ttcacctaga tccttttcac gtagaaagcc agtccgcaga
     8161 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     8221 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     8281 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     8341 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     8401 caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     8461 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     8521 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     8581 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     8641 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     8701 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     8761 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     8821 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     8881 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     8941 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     9001 atgcatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     9061 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     9121 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     9181 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     9241 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     9301 caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     9361 ctaaatacat tcaaatatgt atccgctcat gagattatca aaaaggatct tcacctagat
     9421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     9481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     9541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     9601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     9661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     9721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    10021 cttttctgtg actggtgaat actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    10081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    10141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    10201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    10261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    10321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    10381 tcagggttat tgtctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    10441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    10501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    10561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    10621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    10681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    10741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    10801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    10861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    10921 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    10981 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    11041 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    11101 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    11161 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    11221 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    11281 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
    11341 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
    11401 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    11461 ccatgattac gccaagctca gaattaaccc tcactaaagg g
//
