LOCUS       sequence-448466-       13647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral introduction of BFP reporter (Point mutation H66 in mEGFP
            results in BFP expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13647)
  AUTHORS   Richardson CD, Ray GJ, DeWitt MA, Curie GL, Corn JE
  TITLE     Enhancing homology-directed genome editing by catalytically active 
            and inactive CRISPR-Cas9 using asymmetric donor DNA.
  JOURNAL   Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481.
  PUBMED    26789497
REFERENCE   2  (bases 1 to 13647)
  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: "Nat 
            Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481."
COMMENT     Sequence Label: BFP dest clone
FEATURES             Location/Qualifiers
     source          1..13647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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    2859..2975
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    3009..3033
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3164..4342
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3395..4333
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        4359..4377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4428..4437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4434..4436,4437..4439,4440..5153)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(5168..5267)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5379..5398)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5390..6199
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5457..5476)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6289..6877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      7045..7088
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7220..7247
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     7255..7278
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    7297..7396
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7527..8705
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7758..8696
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        8722..8740
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      8791..8800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8797..8799,8800..8802,8803..9516)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(9531..9555)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    9586..10174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9639..9659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10057..10068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10233..10253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10238..10567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10418..10553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        10615..10662
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10681..11079
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             11223..11456
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11528..11649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11565..11584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11619..11638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11689..11824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(11845..11863)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11873..11890)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11873..11889)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11882..11904)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      12031..12486
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12118..12137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12328..12349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12512..12616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12617..12685,12686..13477)
                     /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     complement(12835..12854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaattcaaa attttatcgt ccagtgtggt ggaattctgc
     3001 agatatcaac aagtttgtac aaaaaagcag gctccaccat gggaaccgac attgattatt
     3061 gactaggctt ttgcaaaaag ctttgcaaag atggataaag ttttaaacag agaggaatct
     3121 ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca
     3181 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     3241 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3301 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3361 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     3421 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     3481 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     3541 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     3601 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     3661 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     3721 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     3781 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     3841 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     3901 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     3961 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     4021 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     4081 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     4141 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     4201 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     4261 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     4321 tcttccattt caggtgtcgt gaggaattag cttggtacta atacgactca ctatagggag
     4381 acccaagctg gctaggtaag cttggtaccg agctcggatc cggtaccgcc accatggtga
     4441 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4501 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4561 tgaccctgaa gttcatctgc accaccggca agctgccggt gccctggccc accctcgtga
     4621 ccaccctgac ccatggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4681 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4741 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4801 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4861 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4921 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4981 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5041 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5101 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taactcgaga
     5161 tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     5221 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
     5281 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     5341 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     5401 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     5461 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     5521 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     5581 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     5641 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     5701 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     5761 gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     5821 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     5881 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     5941 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     6001 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     6061 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     6121 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     6181 gctcgatgag tttttctaat cagaattggt taattggttg taacattatt cagattgggc
     6241 cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6421 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6961 ttctgtggat aaccgtatta ccgctagcat ggatctcggg gacgtctaac tactaagcga
     7021 gagtagggaa ctgccaggca tcaaataaaa cgaaaggccc agtcttccga ctgagccttt
     7081 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     7141 gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     7201 gccaggcatc aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     7261 actcttcctg ttagttagtt acttaagctc gggccccaaa taatgatttt attttgactg
     7321 atagtgacct gttcgttgca acaaattgat aagcaatgct tttttataat gccaactttg
     7381 tacaaaaaag caggctccac catgggaacc gacattgatt attgactagg cttttgcaaa
     7441 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     7501 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     7561 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     7621 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     7681 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     7741 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     7801 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     7861 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     7921 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     7981 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     8041 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     8101 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     8161 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     8221 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     8281 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     8341 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     8401 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     8461 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     8521 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     8581 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     8641 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     8701 cgtgaggaat tagcttggta ctaatacgac tcactatagg gagacccaag ctggctaggt
     8761 aagcttggta ccgagctcgg atccggtacc gccaccatgg tgagcaaggg cgaggagctg
     8821 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     8881 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     8941 tgcaccaccg gcaagctgcc ggtgccctgg cccaccctcg tgaccaccct gacccatggc
     9001 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     9061 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     9121 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     9181 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     9241 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     9301 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     9361 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     9421 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     9481 gggatcactc tcggcatgga cgagctgtac aagtaactcg agatatctag acccagcttt
     9541 cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgacaatca acctctggat
     9601 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     9661 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     9721 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     9781 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     9841 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     9901 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     9961 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
    10021 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
    10081 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
    10141 acgagtcgga tctccctttg ggccgcctcc ccgcctggac ctgcaggatt ctgcagatat
    10201 ccggttagta atgagtttgg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
    10261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    10321 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    10381 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
    10441 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
    10501 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
    10561 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
    10621 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
    10681 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
    10741 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
    10801 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
    10861 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
    10921 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
    10981 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
    11041 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaaa cgcgtcgccc gccccacgac
    11101 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat ctcgagggcc cggtaccttt
    11161 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
    11221 actggaaggg ctagctcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
    11281 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    11341 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    11401 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
    11461 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
    11521 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    11581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    11641 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
    11701 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
    11761 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
    11821 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
    11881 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
    11941 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
    12001 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
    12061 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    12121 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    12181 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    12241 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    12301 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    12361 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    12421 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
    12481 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
    12541 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
    12601 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
    12661 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
    12721 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
    12781 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
    12841 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
    12901 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
    12961 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
    13021 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
    13081 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
    13141 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
    13201 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
    13261 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
    13321 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
    13381 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
    13441 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
    13501 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
    13561 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
    13621 ccccgtagaa aagatcaaag gatcttc
//
