LOCUS       sequence-313967-       12102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lenti sgRNA cloning backbone with MS2 loops and EF1a-MCP-KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12102)
  AUTHORS   Li K, Liu Y, Cao H, Zhang Y, Gu Z, Liu X, Yu A, Kaphle P, Dickerson 
            KE, Ni M, Xu J
  TITLE     Interrogation of enhancer function by enhancer-targeting CRISPR 
            epigenetic editing.
  JOURNAL   Nat Commun. 2020 Jan 24;11(1):485. doi: 10.1038/s41467-020-14362-5.
  PUBMED    31980609
REFERENCE   2  (bases 1 to 12102)
  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 
            Commun."; date: "2020-01-24"; volume: "11(1):485. doi"; pages: " 
            10.1038/s41467-020-14362-5"
COMMENT     Sequence Label: Lenti_sgRNA(MS2)_MCP-KRAB-IRES-zsGreen1
FEATURES             Location/Qualifiers
     source          1..12102
                     /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"
     CDS             1204..1224
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1621..1815
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             1882..1902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1944..2516
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1967..2518
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2111..2128)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             2518..3213
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     primer_bind     3277..3296
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3594..3627)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3690..4278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3743..3763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4161..4172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4281..4297)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4282..4298)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4803..4983
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5009..5026)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5015..5239
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5285..5713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5372..5391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5582..5603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5722..5742)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5727..6056
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5907..6042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5969..5988
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6104..6151
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6170..6544
                     /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    6674..6795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6711..6730)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6765..6784
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6844..6860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6844..6860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6857..6879)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6868..6884
                     /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(6892..6898,6899..6916,6917..6922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6937..6958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7075..7092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7246..7834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7326..7345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8005..8796,8797..8865))
                     /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     8628..8647
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8866..8970)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9045..9064)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9236..9615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9617..9815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9770..9790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             9833..10013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10060..10185
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10678..10911
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11096..11140
                     /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    11438..11555
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        11605..11853
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     11605..11625
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     11776..11795
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11891..11909
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        11961..11979
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
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 aaaatgaagg
      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 gtgtcgtgat gtacaatccg ctagcccacc
     1201 atgcccaaaa agaaaagaaa agtgggtagt atggcttcta actttactca gttcgttctc
     1261 gtcgacaatg gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc
     1321 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt
     1381 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg
     1441 cgttcgtact taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag
     1501 cttattgtta aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc
     1561 gcagcaaact ccggcatcta cggctccgga atggatgcta agtcactgac tgcctggtcc
     1621 cggacactgg tgaccttcaa ggatgtgttt gtggacttca ccagggagga gtggaagctg
     1681 ctggacactg ctcagcagat cctgtacaga aatgtgatgc tggagaacta taagaacctg
     1741 gtttccttgg gttatcagct tactaagcca gatgtgatcc tccggttgga gaagggagaa
     1801 gagccctggc tggtggagag agaaattcac caagagaccc atcctgattc agagactgca
     1861 tttgaaatca aatcatcagt tccgaaaaag aaacgcaaag tttaggaccc gggtcgagca
     1921 tgcatctagg gcggccaatt ccgcccctct ccctcccccc cccctaacgt tactggccga
     1981 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2041 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2101 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     2161 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2221 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2281 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2341 ctcacctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2401 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac
     2461 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2521 gcccagtcca agcacggcct gaccaaggag atgaccatga agtaccgcat ggagggctgc
     2581 gtggacggcc acaagttcgt gatcaccggc gagggcatcg gctacccctt caagggcaag
     2641 caggccatca acctgtgcgt ggtggagggc ggccccttgc ccttcgccga ggacatcttg
     2701 tccgccgcct tcatgtacgg caaccgcgtg ttcaccgagt acccccagga catcgtcgac
     2761 tacttcaaga actcctgccc cgccggctac acctgggacc gctccttcct gttcgaggac
     2821 ggcgccgtgt gcatctgcaa cgccgacatc accgtgagcg tggaggagaa ctgcatgtac
     2881 cacgagtcca agttctacgg cgtgaacttc cccgccgacg gccccgtgat gaagaagatg
     2941 accgacaact gggagccctc ctgcgagaag atcatccccg tgcccaagca gggcatcttg
     3001 aagggcgacg tgagcatgta cctgctgctg aaggacggtg gccgcttgcg ctgccagttc
     3061 gacaccgtgt acaaggccaa gtccgtgccc cgcaagatgc ccgactggca cttcatccag
     3121 cacaagctga cccgcgagga ccgcagcgac gccaagaacc agaagtggca cctgaccgag
     3181 cacgccatcg cctccggctc cgccttgccc tgacctgcag gacggcgagt tcatctacaa
     3241 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3301 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3361 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3421 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3481 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3541 ctccaccggc ggcatggacg agctgtacaa gtaagaattc gtcgagggac ctaataactt
     3601 cgtatagcat acattatacg aagttataca tgtttaaggg ttccggttcc actaggtaca
     3661 attcgaaatt cgatatcaag cttatcggta atcaacctct ggattacaaa atttgtgaaa
     3721 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3781 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3841 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3901 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3961 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4021 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4081 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4141 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4201 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4261 tttgggccgc ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa
     4321 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     4381 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     4441 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     4501 cccaacgaaa acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     4561 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     4621 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     4681 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     4741 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     4801 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     4861 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     4921 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     4981 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5041 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     5101 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     5161 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     5221 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     5281 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5341 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5401 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5461 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5521 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5581 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5641 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5701 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5761 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5881 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     6001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     6061 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     6121 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     6181 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     6241 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     6301 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     6361 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     6421 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     6481 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     6541 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6601 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6661 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6781 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6841 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7021 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7081 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7141 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7201 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7261 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7321 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7381 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7441 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7501 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7561 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7621 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7681 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7741 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7801 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7861 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7921 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7981 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8041 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8101 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8161 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8221 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8281 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8341 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8401 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8461 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8521 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8581 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8641 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8701 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8761 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8821 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8881 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8941 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9001 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     9061 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     9241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     9301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     9361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     9421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     9481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     9541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     9601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     9661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     9721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     9781 ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct
     9841 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     9901 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     9961 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
    10021 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
    10081 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
    10141 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
    10201 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
    10261 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
    10321 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
    10381 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
    10441 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
    10501 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
    10561 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
    10621 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
    10681 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
    10741 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
    10801 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
    10861 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
    10921 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
    10981 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
    11041 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
    11101 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
    11161 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
    11221 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
    11281 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
    11341 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
    11401 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
    11461 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
    11521 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
    11581 atccagtttg gttaattagc tagcgagggc ctatttccca tgattccttc atatttgcat
    11641 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
    11701 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
    11761 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
    11821 gctttatata tcttgtggaa aggacgaaac accggagacg ggataccgtc tctgttttag
    11881 agctaggcca acatgaggat cacccatgtc tgcagggcct agcaagttaa aataaggcta
    11941 gtccgttatc aacttggcca acatgaggat cacccatgtc tgcagggcca agtggcaccg
    12001 agtcggtgct ttttttggat cctgcaaaga tggataaagt tttaaacaga gaggaatctt
    12061 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa tt
//
