LOCUS       sequence-336487-        9836 bp    DNA     circular SYN 14-APR-2025
DEFINITION  2nd generation lentiviral transfer plasmid encoding a U6 promoter 
            expressing a spyCas9 sgRNA targeting B2M and an EF1-a promoter 
            expressing mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9836)
  AUTHORS   Hamilton JR, Tsuchida CA, Nguyen DN, Shy BR, McGarrigle ER, Sandoval
            Espinoza CR, Carr D, Blaeschke F, Marson A, Doudna JA
  TITLE     Targeted delivery of CRISPR-Cas9 and transgenes enables complex 
            immune cell engineering.
  JOURNAL   Cell Rep. 2021 Jun 1;35(9):109207. doi: 
            10.1016/j.celrep.2021.109207.
  PUBMED    34077734
REFERENCE   2  (bases 1 to 9836)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2021-06-1"; volume: "35(9):109207. doi"; pages: " 
            10.1016/j.celrep.2021.109207"
COMMENT     Sequence Label: pCAT005
FEATURES             Location/Qualifiers
     source          1..9836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(83..103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        88..417
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      268..403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     330..349
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        465..512
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             531..905
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1035..1156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1072..1091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1126..1145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1205..1221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1205..1221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1218..1240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1229..1245
                     /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(1253..1259,1260..1277,1278..1283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1298..1319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1436..1453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1607..2195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1687..1706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2366..3157,3158..3226))
                     /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     2989..3008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3227..3331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3406..3425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3591..3970
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3972..4170
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4125..4145
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4188..4368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4415..4540
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5033..5266
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5451..5495
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5793..5910
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5961..6209
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5961..5981
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6132..6151
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        6360..7537
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6591..7528
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7485..7505
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7555..7564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    8287..8875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8340..8360)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8758..8769)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8947..9180
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9206..9223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9212..9436
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(9482..9836,1..74)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9569..9588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9779..9800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
       61 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
      121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
      181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
      361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      421 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
      481 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
      541 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
      601 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
      661 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
      721 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
      781 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
      841 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
      901 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
      961 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     1021 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1141 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     1201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3361 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3421 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3481 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3541 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatatt gacattgatt
     3601 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     3661 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3721 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3781 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3841 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3901 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3961 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     4021 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     4081 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     4141 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt
     4201 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4261 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     4321 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     4381 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     4441 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     4501 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     4561 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     4621 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     4681 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     4741 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     4801 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     4861 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     4921 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     4981 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     5041 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     5101 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     5161 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     5221 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     5281 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     5341 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     5401 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     5461 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     5521 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     5581 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     5641 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     5701 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     5761 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     5821 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     5881 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     5941 gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac
     6001 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
     6061 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     6121 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     6181 tatatatctt gtggaaagga cgaaacaccg gagtagcgcg agcacagcta gtttaagagc
     6241 tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt
     6301 ggcaccgagt cggtgctttt tttgaattcg ctagctaggt cttgaaagga gtgggaattg
     6361 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     6421 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     6481 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     6541 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     6601 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     6661 cttccactgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
     6721 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc
     6781 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
     6841 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
     6901 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
     6961 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
     7021 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
     7081 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
     7141 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
     7201 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
     7261 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
     7321 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
     7381 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
     7441 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
     7501 ggttcaaagt ttttttcttc catttcaggt gtcgtgaacc ggttctagag cgctgccacc
     7561 atggtaagta agggcgaaga ggataatatg gcttctttgc ccgccacaca cgaactgcac
     7621 atattcggat caataaacgg ggtggatttc gatatggtcg gccagggaac aggaaaccct
     7681 aatgacggct acgaagagct caatctcaaa tctaccaaag gggacttaca attctctcca
     7741 tggatcctag tgccacatat tggatatggc ttccatcaat accttccata tccagacggt
     7801 atgagcccct tccaggctgc aatggtggac gggtctggat atcaggttca cagaacaatg
     7861 caatttgaag acggtgcatc ccttacagtt aattacaggt acacatacga aggaagtcac
     7921 atcaaagggg aggcgcaggt gaagggcacc gggttcccag ctgatggacc tgtaatgaca
     7981 aattcactca cagccgccga ttggtgcagg tcaaagaaga cctatcctaa tgacaaaaca
     8041 attatatcca catttaaatg gtcttatacc acaggaaatg gaaagagata cagatctaca
     8101 gctagaacaa cttacacatt tgctaaacca atggcagcca actaccttaa gaatcaacca
     8161 atgtatgtct tcagaaagac tgaactaaaa cacagtaaaa cagaacttaa tttcaaagag
     8221 tggcagaagg ccttcacaga cgttatgggt atggatgaac tgtacaagta aacgcgttaa
     8281 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     8341 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     8401 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     8461 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     8521 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     8581 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     8641 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     8701 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     8761 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     8821 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     8881 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     8941 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     9001 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     9061 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     9121 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     9181 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     9241 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     9301 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     9361 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     9421 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     9481 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     9541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     9601 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     9661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     9721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     9781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttga
//
