LOCUS       sequence-275485-       11950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lenti sgRNA cloning backbone with MS2 loops and EF1a-MCP-VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11950)
  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 11950)
  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-VP64-IRES-zsGreen1
FEATURES             Location/Qualifiers
     source          1..11950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2370..2618
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2370..2390
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2541..2560
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2656..2674
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        2726..2744
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        2868..4046
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3099..4037
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3994..4014
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4071..4091
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4464..4613
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     misc_feature    4659..5231
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4682..5233
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4826..4843)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             5233..5928
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     primer_bind     5992..6011
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(6309..6342)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6405..6993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6458..6478)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6876..6887)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6996..7012)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6997..7013)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7518..7698
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7724..7741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7730..7954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8000..8428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8087..8106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8297..8318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8437..8457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8442..8771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8622..8757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8684..8703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8819..8866
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8885..9259
                     /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    9389..9510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9426..9445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9480..9499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9559..9575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9559..9575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9572..9594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9583..9599
                     /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(9607..9613,9614..9631,9632..9637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9652..9673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9790..9807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9961..10549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10041..10060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10720..11511,11512..11580))
                     /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     11343..11362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11581..11685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11760..11779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttagctagcg agggcctatt tcccatgatt ccttcatatt
     2401 tgcatatacg atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa
     2461 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     2521 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     2581 tcttggcttt atatatcttg tggaaaggac gaaacaccgg agacgggata ccgtctctgt
     2641 tttagagcta ggccaacatg aggatcaccc atgtctgcag ggcctagcaa gttaaaataa
     2701 ggctagtccg ttatcaactt ggccaacatg aggatcaccc atgtctgcag ggccaagtgg
     2761 caccgagtcg gtgctttttt tggatcctgc aaagatggat aaagttttaa acagagagga
     2821 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
     2881 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     2941 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3001 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3061 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     3121 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     3181 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     3241 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3301 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3361 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3421 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3481 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     3541 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     3601 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     3661 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atgaaggacg cggcgctcgg
     3721 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     3781 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     3841 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     3901 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     3961 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4021 tttttcttcc atttcaggtg tcgtgatgta caatccgcta gcccaccatg cccaaaaaga
     4081 aaagaaaagt gggtagtatg gcttctaact ttactcagtt cgttctcgtc gacaatggcg
     4141 gaactggcga cgtgactgtc gccccaagca acttcgctaa cgggatcgct gaatggatca
     4201 gctctaactc gcgttcacag gcttacaaag taacctgtag cgttcgtcag agctctgcgc
     4261 agaatcgcaa atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa
     4321 atatggaact aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg
     4381 caatgcaagg tctcctaaaa gatggaaacc cgattccctc agcaatcgca gcaaactccg
     4441 gcatctacgg ctccggacga gccgatgctc tcgacgattt tgacctcgat atgctgggat
     4501 ccgacgctct cgatgacttc gaccttgaca tgctggggag tgacgctctg gacgacttcg
     4561 atctcgacat gctgggctcc gacgccctgg atgacttcga tctggacatg ctcatcaact
     4621 gacccgggtc gagcatgcat ctagggcggc caattccgcc cctctccctc ccccccccct
     4681 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     4741 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     4801 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     4861 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     4921 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     4981 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     5041 gaaagagtca aatggctcac ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     5101 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     5161 tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     5221 cacgatgata atatggccca gtccaagcac ggcctgacca aggagatgac catgaagtac
     5281 cgcatggagg gctgcgtgga cggccacaag ttcgtgatca ccggcgaggg catcggctac
     5341 cccttcaagg gcaagcaggc catcaacctg tgcgtggtgg agggcggccc cttgcccttc
     5401 gccgaggaca tcttgtccgc cgccttcatg tacggcaacc gcgtgttcac cgagtacccc
     5461 caggacatcg tcgactactt caagaactcc tgccccgccg gctacacctg ggaccgctcc
     5521 ttcctgttcg aggacggcgc cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag
     5581 gagaactgca tgtaccacga gtccaagttc tacggcgtga acttccccgc cgacggcccc
     5641 gtgatgaaga agatgaccga caactgggag ccctcctgcg agaagatcat ccccgtgccc
     5701 aagcagggca tcttgaaggg cgacgtgagc atgtacctgc tgctgaagga cggtggccgc
     5761 ttgcgctgcc agttcgacac cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac
     5821 tggcacttca tccagcacaa gctgacccgc gaggaccgca gcgacgccaa gaaccagaag
     5881 tggcacctga ccgagcacgc catcgcctcc ggctccgcct tgccctgacc tgcaggacgg
     5941 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     6001 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     6061 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     6121 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     6181 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     6241 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag aattcgtcga
     6301 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     6361 gttccactag gtacaattcg aaattcgata tcaagcttat cggtaatcaa cctctggatt
     6421 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6481 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6541 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6601 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6661 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6721 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6781 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     6841 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6901 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6961 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag
     7021 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     7081 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     7141 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     7201 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     7261 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     7321 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     7381 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     7441 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     7501 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7561 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7621 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7681 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     7741 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     7801 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     7861 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     7921 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     7981 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8041 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     8101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8341 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8401 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     8461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     8521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     8581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     8641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     8701 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     8761 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     8821 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     8881 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     8941 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     9001 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     9061 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     9121 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     9181 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     9241 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     9301 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     9361 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     9421 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     9481 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     9541 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     9601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     9661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     9721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     9781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     9841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     9901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     9961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    10081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    10141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    10201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    10261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    10321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    10381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    10441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    10501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    10561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    10621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    10681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    10741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    10801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    10861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    10921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    10981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    11041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    11101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    11161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    11221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    11281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    11341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    11401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    11461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    11521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    11581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    11641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    11701 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
    11761 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
    11821 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
    11881 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
    11941 tatacgcgtt
//
