LOCUS       sequence-474447-       10597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for overexpression KLF4 ORF from a EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10597)
  AUTHORS   Southard KM, Ardy RC, Tang A, O'Sullivan DD, Metzner E, 
            Guruvayurappan K, Norman TM
  TITLE     Comprehensive transcription factor perturbations recapitulate 
            fibroblast transcriptional states.
  JOURNAL   Nat Genet. 2025 Aug 6. doi: 10.1038/s41588-025-02284-1.
  PUBMED    40770575
REFERENCE   2  (bases 1 to 10597)
  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 Genet. 
            2025 Aug 6. doi: 10.1038/s41588-025-02284-1."
COMMENT     Sequence Label: pRCA881 - KLF4 Puro-T2A-GFP (pRCA847 backbone)
FEATURES             Location/Qualifiers
     source          1..10597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2743..3331
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2796..2816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3214..3225)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3403..3636
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3714..3835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3751..3770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3805..3824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3875..4010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4030..4049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4031..4049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4059..4076)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4059..4075)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4068..4090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4217..4672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4514..4535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4803..4871,4872..5663)
                     /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(5021..5040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6576..6593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6710..6731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6746..6751,6752..6769,6770..6776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6800
                     /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     6789..6811
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6808..6824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6808..6824
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6843..6863
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6845..6863
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6891..7117
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7118..7298
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7345..7470
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7963..8196
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8381..8425
                     /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"
     promoter        8656..8866
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        8678..8874
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8725..8860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8906..9502
                     /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(8906..8925)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9402..9422
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9515..9568
                     /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"
     regulatory      9575..9584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9581..9583,9584..9586,9587..10300)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(9581..9583,9584..9586,9587..10297)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9626..9647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9887..9906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10234..10255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10343..10460
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gctagcaccg ccaccatgag
     1201 gcagccacct ggcgagtctg acatggctgt cagcgacgcg ctgctcccat ctttctccac
     1261 gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt caagcaggtg ccccgaataa
     1321 ccgctggcgg gaggagctct cccacatgaa gcgacttccc ccagtgcttc ccggccgccc
     1381 ctatgacctg gcggcggcga ccgtggccac agacctggag agcggcggag ccggtgcggc
     1441 ttgcggcggt agcaacctgg cgcccctacc tcggagagag accgaggagt tcaacgatct
     1501 cctggacctg gactttattc tctccaattc gctgacccat cctccggagt cagtggccgc
     1561 caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg ccgtcgagca gcggccctgc
     1621 cagcgcgccc tccacctgca gcttcaccta tccgatccgg gccgggaacg acccgggcgt
     1681 ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg gagtccgctc cccctccgac
     1741 ggctcccttc aacctggcgg acatcaacga cgtgagcccc tcgggcggct tcgtggccga
     1801 gctcctgcgg ccagaattgg acccggtgta cattccgccg cagcagccgc agccgccagg
     1861 tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg agcgcccctg gcagcgagta
     1921 cggcagcccg tcggtcatca gcgtcagcaa aggcagccct gacggcagcc acccggtggt
     1981 ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc aagatcaagc aggaggcggt
     2041 ctcttcgtgc acccacttgg gcgctggacc ccctctcagc aatggccacc ggccggctgc
     2101 acacgacttc cccctggggc ggcagctccc cagcaggact accccgaccc tgggtcttga
     2161 ggaagtgctg agcagcaggg actgtcaccc tgccctgccg cttcctcccg gcttccatcc
     2221 ccacccgggg cccaattacc catccttcct gcccgatcag atgcagccgc aagtcccgcc
     2281 gctccattac caagagctca tgccacccgg ttcctgcatg ccagaggagc ccaagccaaa
     2341 gaggggaaga cgatcgtggc cccggaaaag gaccgccacc cacacttgtg attacgcggg
     2401 ctgcggcaaa acctacacaa agagttccca tctcaaggca cacctgcgaa cccacacagg
     2461 tgagaaacct taccactgtg actgggacgg ctgtggatgg aaattcgccc gctcagatga
     2521 actgaccagg cactaccgta aacacacggg gcaccgcccg ttccagtgcc aaaaatgcga
     2581 ccgagcattt tccaggtcgg accacctcgc cttacacatg aagaggcatt tttagtaatg
     2641 aactagtccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     2701 cgaccattcc gcgtgaagtg aactcggacg cgttaagtcg acaatcaacc tctggattac
     2761 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2821 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2881 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2941 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3001 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3061 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3121 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3181 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3241 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3301 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     3361 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3421 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     3481 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3541 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3601 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     3661 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     3721 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3781 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3841 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3901 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3961 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     4021 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     4081 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4141 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4201 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6541 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6601 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6661 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6721 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6781 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6841 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     6901 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     6961 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     7021 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     7081 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7141 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7201 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7261 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7321 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7381 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7441 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7501 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7561 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7621 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7681 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7741 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7801 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7861 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7921 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7981 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     8041 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8101 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8161 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8221 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8281 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8341 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8401 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8461 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8521 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8581 cccaaccccg aggggaccca tgcattgcat ctcaattagt cagcaaccag gtgtggaaag
     8641 tccccaggct ccccagcagg cagaagtatg caaagcatgc gtctcaatta gtcagcaacc
     8701 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     8761 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     8821 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     8881 tctagaggta ccaccggagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     8941 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     9001 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     9061 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     9121 cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     9181 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     9241 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     9301 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     9361 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     9421 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     9481 catgacccgc aagcccggtg ccagtggatc gggagagggc agaggaagtc tgctaacatg
     9541 cggtgacgtc gaggagaatc ctggcccacc ggtcgccacc atggtgagca agggcgagga
     9601 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     9661 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     9721 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     9781 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     9841 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     9901 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     9961 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
    10021 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
    10081 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
    10141 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
    10201 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
    10261 cgccgggatc actctcggca tggacgagct gtacaagtaa cgaattctag atcttgagac
    10321 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
    10381 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
    10441 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc
    10501 ggctcgaggc ctgcaggtgc aaagatggat aaagttttaa acagagagga atctttgcag
    10561 ctaatggacc ttctaggtct tgaaaggagt gggaatt
//
