LOCUS       sequence-322106-       10201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for negative control of knockdown expressing a 
            non-target miR against Renilla luciferase, with expression of a Ly6G
            surface marker and a puromycin selection marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10201)
  AUTHORS   van der Donk LEH, van der Spek J, van Duivenvoorde T, Ten Brink MS, 
            Geijtenbeek TBH, Kuijl CP, van Heijst JWJ, Ates LS
  TITLE     An optimized retroviral toolbox for overexpression and genetic 
            perturbation of primary lymphocytes.
  JOURNAL   Biol Open. 2022 Feb 15;11(2):bio059032. doi: 10.1242/bio.059032. 
            Epub 2022 Mar 1.
  PUBMED    35229875
REFERENCE   2  (bases 1 to 10201)
  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: "Biol 
            Open."; date: "2022-02-15"; volume: "11(2):bio059032. doi"; pages: "
            10.1242/bio.059032. Epub 2022 Mar 1"
COMMENT     Sequence Label: pLenti-EF1a-Ly6G-P2A-PuroR-HA-miR30
FEATURES             Location/Qualifiers
     source          1..10201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1066..1299
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1484..1528
                     /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    1826..1943
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1993..3175
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2224..3166
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3123..3143
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3209..3218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3689..4285
                     /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(3689..3708)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4185..4205
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4295..4321
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     ncRNA           4348..4469
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4497..4511
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4539..4665
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    4697..5285
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4750..4770)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5168..5179)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5357..5590
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5616..5633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5622..5846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5892..6320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5979..5998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6189..6210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6329..6349)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6334..6663
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6514..6649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6576..6595
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6711..6758
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6777..7151
                     /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    7281..7402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7318..7337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7372..7391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7451..7467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7451..7467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7464..7486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7475..7491
                     /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(7499..7505,7506..7523,7524..7529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7544..7565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7682..7699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7853..8441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7933..7952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8612..9403,9404..9472))
                     /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     9235..9254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9473..9577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9652..9671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9821..10200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
      961 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1021 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1081 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1141 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1201 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1261 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1321 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     1381 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1441 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1501 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1561 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     1621 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     1681 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     1741 agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca
     1801 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     1861 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     1921 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
     1981 taattaacgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     2041 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     2101 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     2161 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     2221 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     2281 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     2341 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     2401 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     2461 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     2521 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     2581 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     2641 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     2701 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     2761 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     2821 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     2881 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     2941 aggcacctcg attagttctc gatcgagctt ttggagtacg tcgtctttag gttgggggga
     3001 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     3061 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     3121 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaaggat
     3181 ccggtcgtgc ggccgcacgg aaacgcgtgc caccatggac acctgtcata tcgccaagag
     3241 ctgcgtgctg atcctgctgg tggttctgct gtgtgccgag cgagcacagg gactggagtg
     3301 ctacaactgt atcggcgtgc cacctgagac aagctgcaac accaccacct gtcctttcag
     3361 cgacggcttc tgtgtggccc tggaaatcga agtgatcgtg gacagccacc gcagcaaagt
     3421 gaagtccaac ctgtgcctgc ctatctgccc caccacactg gacaacaccg agatcacagg
     3481 caacgccgtg aacgtgaaaa cctactgctg caaagaggac ctctgcaacg ccgctgttcc
     3541 aacaggcgga agctcttgga caatggctgg cgtgctgctg ttcagcctgg tgtctgttct
     3601 gctgcagacc ttcctgggca gcggcgtcga cgccacaaac ttcagcctgc tgagacaggc
     3661 tggcgacgtg gaagagaatc ctggacctat gaccgagtac aagcccacgg tgcgcctcgc
     3721 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     3781 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     3841 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     3901 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     3961 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4021 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4081 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4141 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4201 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4261 gtgcatgacc cgcaagcccg gtgccggcag cggctaccct tacgatgtgc ctgattacgc
     4321 ctgacctgca ggtttctgaa gctttgtttg aatgaggctt cagtacttta cagaatcgtt
     4381 gcctgcacat cttggaaaca cttgctggga ttacttcttc aggttaaccc aacagaaggc
     4441 tcgagaaggt atattgctgt tgacagtgag cgaaagacaa tgagtacaca gcaatagtga
     4501 agccacagat gtattgctgt gtactcattg tcttctgcct actgcctcgg aattcaaggg
     4561 gctactttag gagcaattat cttgtttact aaaactgaat accttgctat ctctttgata
     4621 catttttaca aagctgaatt aaaatggtat aaattaaatc actttatcga tacggaaacg
     4681 cgcgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4741 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4801 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4861 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4921 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4981 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5041 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5101 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5161 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5221 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5281 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5341 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     5401 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5461 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5521 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5581 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     5641 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5701 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5761 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5821 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     5881 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6241 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6301 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     6361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     6421 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6481 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6541 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6601 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6661 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     6721 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     6781 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     6841 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     6901 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     6961 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     7021 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     7081 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     7141 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7201 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7261 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     7321 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     7381 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     7441 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7501 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7561 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7621 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7681 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7741 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7801 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8281 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9601 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     9661 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     9721 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     9781 gaatctgctt agggttaggc gttttgcgct gcttcgcgtt gacattgatt attgactagt
     9841 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     9901 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     9961 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
    10021 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
    10081 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
    10141 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
    10201 g
//
