LOCUS       sequence-308792-       11729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock IL13-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 11729)
  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 11729)
  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 IL13-CAR
FEATURES             Location/Qualifiers
     source          1..11729
                     /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    5091..5679
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5144..5164)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5562..5573)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5711..6187
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5910..5929)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6297..6316)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6362..6417
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6416..6435)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6778..6794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6802..6818
                     /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(6826..6832,6833..6850,6851..6856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6871..6892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6966..7802
                     /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(7836..7855)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7837..7855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7862..7879)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7862..7878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7871..7893)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8016..8318
                     /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     8197..8216
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8721..8740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9335..9354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9715..10506,10507..10575))
                     /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     10338..10357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10699..11287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11188..11207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11441..11458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11575..11596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11611..11616,11617..11634,11635..11641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11649..11665
                     /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     11673..11689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11710..11728
                     /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 ctgatcccag gccctgtgcc tccctctaca gccctcaggt
     3661 acctcattga ggagctggtc aacatcaccc agaaccagaa ggctccgctc tgcaatggca
     3721 gcatggtatg gagcatcaac ctgacagctg gcatgtactg tgcagccctg gaatccctga
     3781 tcaacgtgtc aggctgcagt gccatcgaga agacccagag gatgctgagc ggattctgcc
     3841 cgcacaaggt ctcagctggg cagttttcca gcttgcatgt ccgagacacc aaaatcgagg
     3901 tggcccagtt tgtaaaggac ctgctcttac atttaaagaa actttttcgc gagggacggt
     3961 tcaacgagtc caaatatggt cccccatgcc caccatgccc agcacctgag ttcctggggg
     4021 gaccatcagt cttcctgttc cccccaaaac ccaaggacac tctcatgatc tcccggaccc
     4081 ctgaggtcac gtgcgtggtg gtggacgtga gccaggaaga ccccgaggtc cagttcaact
     4141 ggtacgtgga tggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagttca
     4201 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaacggca
     4261 aggagtacaa gtgcaaggtc tccaacaaag gcctcccgtc ctccatcgag aaaaccatct
     4321 ccaaagccaa agggcagccc cgagagccac aggtgtacac cctgccccca tcccaggagg
     4381 agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctac cccagcgaca
     4441 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg
     4501 tgctggactc cgacggctcc ttcttcctct acagcaggct aaccgtggac aagagcaggt
     4561 ggcaggaggg gaatgtcttc tcatgctccg tgatgcatga ggctctgcac aaccactaca
     4621 cacagaagag cctctccctg tccctaggta aaatggccct gattgtgctg gggggcgtcg
     4681 ccggcctcct gcttttcatt gggctaggca tcttcttcag agtgaagttc agcaggagcg
     4741 cagacgcccc cgcgtaccag cagggccaga accagcagct ctataacgag ctcaatctag
     4801 gacgaagaga ggagtacgat gttttggaca agagacgtgg ccgggaccct gagatggggg
     4861 gaaagccgag aaggaagaac cctcaggaag gcctgtacaa tgaactgcag aaagataaga
     4921 tggcggaggc ctacagtgag attgggatga aaggcgagcg ccggaggggc aaggggcacg
     4981 atggccttta ccagggtctc agtacagcca ccaaggacac ctacgacgcc cttcacatgc
     5041 aggccctgcc ccctcgctga acgcgtgaat tcgatatcaa gcttatcgat aatcaacctc
     5101 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5161 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5221 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5281 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5341 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5401 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5461 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     5521 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5581 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5641 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     5701 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     5761 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     5821 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     5881 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     5941 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     6001 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     6061 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     6121 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     6181 tgtccttctg attttgtagg taaccaggcg cgtgaattca ctcctcaggt gcaggctgcc
     6241 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     6301 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     6361 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     6421 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     6481 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     6541 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     6601 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     6661 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     6721 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     6781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcg acagtactaa
     6961 gctttactag ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta
     7021 gtctcctgat attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca
     7081 cacccccatt tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat
     7141 ggagccagag aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat
     7201 gcgtgacctg cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg
     7261 ctctgacgcg gctgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc
     7321 ccaagaggag aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact
     7381 ttggctcttc acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct
     7441 gtgagataag gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc
     7501 cgtgtggaaa actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg
     7561 cctctactcc ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg
     7621 ggacatattc ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct
     7681 gggtttgagt ccttggcaag cccaggagag gcgctcaggc ttccctgtcc cccttcctcg
     7741 tccaccatct catgcccctg gctctcctgc cccttcccta caggggttcc tggctctgct
     7801 ctaagggcga attcgcggcc gctaaattca attcgcccta tagtgagtcg tattacaatt
     7861 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     7921 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     7981 gcccttccca acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa
     8041 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc
     8101 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
     8161 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     8221 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     8281 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa
     8341 aaaggatctt cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac
     8401 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
     8461 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
     8521 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     8581 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc
     8641 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     8701 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     8761 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     8821 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     8881 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     8941 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     9001 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     9061 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     9121 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     9181 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat gcatggcgat
     9241 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     9301 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     9361 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     9421 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac
     9481 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg
     9541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     9601 aaatatgtat ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    10021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    10081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    10141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    10201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    10261 tggtgaatac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    10321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    10381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    10441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    10501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    10561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    10621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
    10681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
    10741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
    10801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
    10861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
    10921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
    10981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
    11041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
    11101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
    11161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
    11221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
    11281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
    11341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
    11401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
    11461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
    11521 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
    11581 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
    11641 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
    11701 caagctcaga attaaccctc actaaaggg
//
