LOCUS       sequence-272646-       10963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For simple determination of the multiplicity of infection (MOI) of a
            lentiviral CRISPR library by checking EGFP expression (still 
            allowing for puro selection)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10963)
  AUTHORS   Panda SK, Wigerblad G, Jiang L, Jimenez-Andrade Y, Iyer VS, Shen Y, 
            Boddul SV, Guerreiro-Cacais AO, Raposo B, Kasza Z, Wermeling F
  TITLE     IL-4 controls activated neutrophil FcgammaR2b expression and 
            migration into inflamed joints.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jan 24. pii: 1914186117. doi: 
            10.1073/pnas.1914186117.
  PUBMED    31980518
REFERENCE   2  (bases 1 to 10963)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Jan 24. pii: 1914186117. doi: 
            10.1073/pnas.1914186117."
COMMENT     Sequence Label: LentiGuide Puro-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..10963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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        1486..1734
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1486..1506
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1657..1676
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3616..3691
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3747..3864
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4003..5181
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4234..5172
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5129..5149
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5194..5790
                     /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(5194..5213)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5690..5710
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5800..5856
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(5857..5859,5860..6573)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5857..5859,5860..6570)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5899..5920)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6160..6179)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6507..6528
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6589..7177
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6642..6662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7060..7071)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7249..7482
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7560..7681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7597..7616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7651..7670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7721..7856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7876..7895)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7877..7895)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7905..7922)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7905..7921)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7914..7936)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8063..8518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8150..8169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8360..8381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8544..8648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8649..8717,8718..9509)
                     /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(8867..8886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9680..10268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10169..10188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10422..10439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10556..10577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10592..10597,10598..10615,10616..10622)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10630..10646
                     /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     10635..10657
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10654..10670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10654..10670
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10689..10709
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10691..10709
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10737..10963
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagagaggg cctatttccc
     1501 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tagaattaat
     1561 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     1621 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     1681 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggagac
     1741 ggttgtaaat gagcacacaa aatacacatg ctaaaatatt atattctatg acctttataa
     1801 aatcaaccaa aatcttcttt ttaataactt tagtatcaat aattagaatt tttatgttcc
     1861 tttttgcaaa cttttaataa aaatgagcaa aataaaaaaa cgctagtttt agtaactcgc
     1921 gttgttttct tcacctttaa taatagctac tccaccactt gttcctaagc ggtcagctcc
     1981 tgcttcaatc attttttgag catcttcaaa tgttctaact ccaccagctg ctttaactaa
     2041 agcattgtct ttaacaactg acttcattag tttaacatct tcaaatgttg cacctgattt
     2101 tgaaaatcct gttgatgttt taacaaattc taatccagct tcaacagcta tttcacaagc
     2161 tttcatgatt tcttcttttg ttaataaaca attttccata atacatttaa caacatgtga
     2221 tccagctgct ttttttacag ctttcatgtc ttctaaaact aattcataat ttttgtcttt
     2281 taatgcacca atatttaata ccatatcaat ttctgttgca ccatctttaa ttgcttcaga
     2341 aacttcgaat gcttttgtag ctgttgtgca tgcacctaga ggaaaaccta caacatttgt
     2401 tattcctaca tttgtgcctt ttaataattc tttacaatag cttgttcaat atgaattaac
     2461 acaaactgtt gcaaaatcaa attcaattgc ttcatcacat aattgtttaa tttcagcttt
     2521 cgtagcatct tgttttaata atgtgtgatc tatatatttg tttagtttca ttttttctcc
     2581 tatatattca tttttaattt taattcttta ataatttcgt ctactttaac tttagcgttt
     2641 tgaacagatt caccaacacc tataaaataa atttttagtt taggttcagt tccacttggg
     2701 cgaacagcaa atcatgactt atcttctaaa taaaatttta gtaagtcttg tcctggcata
     2761 ttatacattc catcgatgta gtcttcaaca ttaacaactt taagtccagc aatttgagtt
     2821 aagggtgttg ctctcaatga tttcattaat ggttcaattt ttaatttctt ttcttctggt
     2881 ttaaaattca agtttaaagt gaaagtgtaa tatgcaccca tttctttaaa taaatcttct
     2941 aaatagtcta ctaatgtttt attttgtttt ttataaaatc aagcagcctc tgctattaat
     3001 atagaagctt gtattccatc tttatctcta gctgagtcat caattacata tccataactt
     3061 tcttcataag caaaaacaaa atttaatccg ttatcttctt ctttagcaat ttctctaccc
     3121 attcatttaa atccagttaa agtttttaca atattaactc catatttttc atgagcgatt
     3181 ctatcaccca aatcacttgt tacaaaactt gaatatagag ccggattttt tggaatgcta
     3241 tttaagcgtt ttagatttga taattttcaa tcaattaaaa ttggtcctgt ttgatttcca
     3301 tctaatctta caaaatgacc atcatgtttt attgccattc caaatctgtc agcatctggg
     3361 tcattcataa taataatatc tgcatcatgt ttaataccat attcaagcgg tatttttcat
     3421 gcaggatcaa attctggatt tggatttaca acatttttaa atgtttcatc ttcaaatgca
     3481 tgctcttcaa cctcaataac gttatatcct gattcacgta atatttttgg ggtaaattta
     3541 gttcctgttc cattaactgc gctaaaaata atttttaaat cttttttagc ttcttgctct
     3601 tttttgtacg tctctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     3661 tcaacttgaa aaagtggcac cgagtcggtg cttttttaag cttggcgtaa ctagatcttg
     3721 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     3781 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     3841 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     3901 cgccggctcg agggggcccg ggtgcaaaga tggataaagt tttaaacaga gaggaatctt
     3961 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     4021 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4081 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4141 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4201 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     4261 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
     4321 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
     4381 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
     4441 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
     4501 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
     4561 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
     4621 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
     4681 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
     4741 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
     4801 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
     4861 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
     4921 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
     4981 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     5041 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     5101 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     5161 cttccatttc aggtgtcgtg acgtacggcc accatgaccg agtacaagcc cacggtgcgc
     5221 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5281 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     5341 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5401 ggcgccgccg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5461 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5521 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga
     5581 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5641 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5701 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5761 acctggtgca tgacccgcaa gcccggtgcc ggatccggcg caacaaactt ctctctgctg
     5821 aaacaagccg gagatgtcga agagaatcct ggaccggtga gcaagggcga ggagctgttc
     5881 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5941 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     6001 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     6061 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     6121 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     6181 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     6241 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     6301 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     6361 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     6421 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     6481 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     6541 atcactctcg gcatggacga gctgtacaag taaacgcgtt aagtcgacaa tcaacctctg
     6601 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6661 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6721 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6781 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6841 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6901 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6961 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     7021 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     7081 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     7141 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     7201 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     7261 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     7321 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7381 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7441 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat
     7501 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     7561 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7621 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7681 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     7741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     7801 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     7861 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     7921 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     7981 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     8041 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     8101 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     8161 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     8221 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     8281 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     8341 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     8401 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     8461 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     8521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     8581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     8641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     8701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     8761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     8821 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     8881 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     8941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     9001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     9061 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     9121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     9181 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     9241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     9301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     9361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     9421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     9481 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     9541 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     9601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     9661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     9721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     9781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     9841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     9901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     9961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    10021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    10081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    10141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    10201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    10261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    10321 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
    10381 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
    10441 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
    10501 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
    10561 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
    10621 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
    10681 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
    10741 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
    10801 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
    10861 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
    10921 ttgcagagat attgtattta agtgcctagc tcgatacata aac
//
