LOCUS       sequence-308789-       11858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock CLTX-NKG2D-2B4-CD3z 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 11858)
  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 11858)
  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-NKG2D-2B4-CD3z CAR
FEATURES             Location/Qualifiers
     source          1..11858
                     /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"
     CDS             complement(4740..4751)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    5220..5808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5273..5293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5691..5702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5840..6316
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6039..6058)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6426..6445)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6491..6546
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6545..6564)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6907..6923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6931..6947
                     /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(6955..6961,6962..6979,6980..6985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7000..7021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7095..7931
                     /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(7965..7984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7966..7984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7991..8008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7991..8007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8000..8022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8145..8447
                     /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     8326..8345
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8850..8869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9464..9483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9844..10635,10636..10704))
                     /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     10467..10486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10828..11416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11317..11336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11570..11587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11704..11725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11740..11745,11746..11763,11764..11770)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11778..11794
                     /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     11802..11818
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11839..11857
                     /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 ccaggcgggg
     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 cagaggccag 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 taaaccattt tttttctgct
     4441 gcttcatcgc tgtagccatg ggaatccgtt tcattattat ggtaacatgg aggagaaaga
     4501 ggaaggagaa gcagtcagag accagtccca aggaattttt gacaatttac gaagatgtca
     4561 aggatctgaa aaccaggaga aatcacgagc aggagcagac ttttcctgga ggggggagca
     4621 ccatctactc tatgatccag tcccagtctt ctgctcccac gtcacaagaa cctgcatata
     4681 cattatattc attaattcag ccttccagga agtctggatc caggaagagg aaccacagcc
     4741 cttccttcaa tagcactatc tatgaagtga ttggaaagag tcaacctaaa gcccagaacc
     4801 ctgctcgatt gagccgcaaa gagctggaga actttgatgt ttattccaga gtgaagttca
     4861 gcaggagcgc agacgccccc gcgtaccagc agggccagaa ccagcagctc tataacgagc
     4921 tcaatctagg acgaagagag gagtacgatg ttttggacaa gagacgtggc cgggaccctg
     4981 agatgggggg aaagccgaga aggaagaacc ctcaggaagg cctgtacaat gaactgcaga
     5041 aagataagat ggcggaggcc tacagtgaga ttgggatgaa aggcgagcgc cggaggggca
     5101 aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc tacgacgccc
     5161 ttcacatgca ggccctgccc cctcgctgaa cgcgtgaatt cgatatcaag cttatcgata
     5221 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5281 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5341 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5401 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5461 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5521 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5581 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5641 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5701 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5761 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     5821 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     5881 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     5941 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     6001 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     6061 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     6121 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     6181 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     6241 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     6301 tcccttccct gtccttctga ttttgtaggt aaccaggcgc gtgaattcac tcctcaggtg
     6361 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     6421 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     6481 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     6541 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     6601 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     6661 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     6721 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     6781 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     6841 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     6901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga
     7081 cagtactaag ctttactagg gacaggattg gtgacagaaa agccccatcc ttaggcctcc
     7141 tccttcctag tctcctgata ttgggtctaa cccccacctc ctgttaggca gattccttat
     7201 ctggtgacac acccccattt cctggagcca tctctctcct tgccagaacc tctaaggttt
     7261 gcttacgatg gagccagaga ggatcctggg agggagagct tggcaggggg tgggagggaa
     7321 gggggggatg cgtgacctgc ccggttctca gtggccaccc tgcgctaccc tctcccagaa
     7381 cctgagctgc tctgacgcgg ctgtctggtg cgtttcactg atcctggtgc tgcagcttcc
     7441 ttacacttcc caagaggaga agcagtttgg aaaaacaaaa tcagaataag ttggtcctga
     7501 gttctaactt tggctcttca cctttctagt ccccaattta tattgttcct ccgtgcgtca
     7561 gttttacctg tgagataagg ccagtagcca gccccgtcct ggcagggctg tggtgaggag
     7621 gggggtgtcc gtgtggaaaa ctccctttgt gagaatggtg cgtcctaggt gttcaccagg
     7681 tcgtggccgc ctctactccc tttctctttc tccatccttc tttccttaaa gagtccccag
     7741 tgctatctgg gacatattcc tccgcccaga gcagggtccc gcttccctaa ggccctgctc
     7801 tgggcttctg ggtttgagtc cttggcaagc ccaggagagg cgctcaggct tccctgtccc
     7861 ccttcctcgt ccaccatctc atgcccctgg ctctcctgcc ccttccctac aggggttcct
     7921 ggctctgctc taagggcgaa ttcgcggccg ctaaattcaa ttcgccctat agtgagtcgt
     7981 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     8041 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     8101 gcaccgatcg cccttcccaa cagttgcgca gcctatacgt acggcagttt aaggtttaca
     8161 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
     8221 cgccggggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
     8281 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
     8341 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
     8401 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggcatga
     8461 gattatcaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac
     8521 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     8581 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     8641 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     8701 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     8761 gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     8821 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     8881 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     8941 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     9001 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     9061 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     9121 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     9181 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     9241 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     9301 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     9361 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     9421 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     9481 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     9541 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattatt
     9601 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     9661 cgcatcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     9721 aatacattca aatatgtatc cgctcatgag attatcaaaa aggatcttca cctagatcct
     9781 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     9841 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     9901 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     9961 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    10021 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    10081 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    10141 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    10201 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    10261 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    10321 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    10381 ttctgtgact ggtgaatact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    10441 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    10501 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    10561 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    10621 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    10681 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    10741 gggttattgt ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
    10801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
    10861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
    10921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
    10981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
    11041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
    11101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
    11161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
    11221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
    11281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    11341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    11401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
    11461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
    11521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
    11581 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
    11641 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    11701 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    11761 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
    11821 tgattacgcc aagctcagaa ttaaccctca ctaaaggg
//
