LOCUS       sequence-142766-        8297 bp    DNA     circular SYN 14-APR-2025
DEFINITION  sgRNA1-2XboxB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8297)
  AUTHORS   Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
  TITLE     Multiplexed labeling of genomic loci with dCas9 and engineered 
            sgRNAs using CRISPRainbow.
  JOURNAL   Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
  PUBMED    27088723
REFERENCE   2  (bases 1 to 8297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526."
COMMENT     Sequence Label: pLH-sgRNA1-2XboxB
FEATURES             Location/Qualifiers
     source          1..8297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             597..1622
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             597..1622
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(1646..1662)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1647..1663)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1745..1978
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2050..2171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2087..2106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2141..2160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2211..2346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2366..2385)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2367..2385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2395..2412)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2395..2411)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2404..2426)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2553..3008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2640..2659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2850..2871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3034..3138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3139..3207,3208..3999)
                     /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(3357..3376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4170..4758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4659..4678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4912..4929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5046..5067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5082..5087,5088..5105,5106..5112)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5120..5136
                     /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     5125..5147
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5144..5160
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5144..5160
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5179..5199
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5181..5199
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5227..5453
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5454..5634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5681..5806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6299..6532
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6717..6761
                     /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        7059..7307
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7059..7079
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7230..7249
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             7648..7953
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     7832..7851
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    7986..8004
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    8041..8059
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     misc_feature    8128..8245
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8294..8297,1..507)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg atcaagcttg ccagctgggg
      541 cgccctctgg taaggttggg aagccctgca gatctgatcc cggggggcaa tgagatatga
      601 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
      661 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
      721 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
      781 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
      841 aatttagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
      901 atctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
      961 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     1021 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     1081 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     1141 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     1201 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     1261 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     1321 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     1381 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     1441 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     1501 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     1561 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     1621 agagtagatg ccgaccggga tctatcgata ccgtcgacct cgaggggggg cccggtacct
     1681 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     1741 ggactggaag ggctaattca ctcccaacga agaaaagatc tgctttttgc ttgtactggg
     1801 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     1861 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     1921 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     1981 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     2041 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2101 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2161 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     2221 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2281 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2341 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     2401 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     2461 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2521 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2701 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2761 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2881 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3001 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3121 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3181 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3361 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3661 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3961 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4021 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4141 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4801 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4861 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4921 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4981 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5041 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5101 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5161 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
     5221 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc
     5281 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt
     5341 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg
     5401 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc
     5461 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5521 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5581 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     5641 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     5701 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     5761 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     5821 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     5881 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     5941 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     6001 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     6061 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     6121 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     6181 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     6241 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     6301 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     6361 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     6421 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     6481 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     6541 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     6601 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     6661 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     6721 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     6781 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     6841 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     6901 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     6961 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc
     7021 gatcacgaga ctagcctcga gcggccgccc ccttcaccga gggcctattt cccatgattc
     7081 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     7141 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     7201 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     7261 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggt gtcttcgagg
     7321 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     7381 atatactgat atgtataccc gaagtatgtc aaaaagaggt gtgctatgaa gcagcgtatt
     7441 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     7501 ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     7561 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     7621 ttgctgacga gaacagggac tggtgaaatg cagtttaagg tttacaccta taaaagagag
     7681 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     7741 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     7801 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     7861 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     7921 aacgccatta acctgatgtt ctggggaata taaaccctgc aggacgaaga cccgtttgag
     7981 agctagggcc ctgaagaagg gccctagcaa gttcaaataa ggctagtccg ttatcaactt
     8041 gggccctgaa gaagggccca agtggcaccg agtcggtgct ttttttgaat tctcgacctc
     8101 gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     8161 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     8221 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg
     8281 gccgcggctc gaggggg
//
