LOCUS       sequence-331359-       11243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to co-express an sasgRNA with NLS-mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11243)
  AUTHORS   Wang S, Matsumoto K, Lish SR, Cartagena-Rivera AX, Yamada KM
  TITLE     Budding epithelial morphogenesis driven by cell-matrix versus 
            cell-cell adhesion.
  JOURNAL   Cell. 2021 Jul 8;184(14):3702-3716.e30. doi: 
            10.1016/j.cell.2021.05.015. Epub 2021 Jun 15.
  PUBMED    34133940
REFERENCE   2  (bases 1 to 11243)
  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"; doi: 
            "10.1016/j.cell.2021.05.015"; journalName: "Cell"; date: "2021-07-8-
            8"; volume: "184"; issue: "14"; pages: "3702-3716.e30"
COMMENT     Sequence Label: 
            pW214-lenti-sasgRNA-Esp3I-2kb-filler-pEF1s-NLS-mScarlet-I-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..11243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1990..2010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2161..2180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4120..4195
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        4257..4468
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4496..4543
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4544..5236
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(4674..4692)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4942..4961
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5246..5302
                     /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             5303..5701
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5717..6305
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5770..5790)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6188..6199)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6377..6610
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6636..6653)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6642..6866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6912..7340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6999..7018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7209..7230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7349..7369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7354..7683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7534..7669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7596..7615
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7731..7778
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7797..8171
                     /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    8301..8422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8338..8357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8392..8411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8471..8487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8471..8487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8484..8506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8495..8511
                     /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(8519..8525,8526..8543,8544..8549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8564..8585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8702..8719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8873..9461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8953..8972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9632..10423,10424..10492))
                     /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     10255..10274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10493..10597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10672..10691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10863..11242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa
     2281 tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat
     2341 caataattag aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa
     2401 aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc
     2461 acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct
     2521 aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac
     2581 atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc
     2641 agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc
     2701 cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa
     2761 aactaattca taatttttgt cttttaatgc accaatattt aataccatat caatttctgt
     2821 tgcaccatct ttaattgctt cagaaacttc gaatgctttt gtagctgttg tgcatgcacc
     2881 tagaggaaaa cctacaacat ttgttattcc tacatttgtg ccttttaata attctttaca
     2941 atagcttgtt caatatgaat taacacaaac tgttgcaaaa tcaaattcaa ttgcttcatc
     3001 acataattgt ttaatttcag ctttcgtagc atcttgtttt aataatgtgt gatctatata
     3061 tttgtttagt ttcatttttt ctcctatata ttcattttta attttaattc tttaataatt
     3121 tcgtctactt taactttagc gttttgaaca gattcaccaa cacctataaa ataaattttt
     3181 agtttaggtt cagttccact tgggcgaaca gcaaatcatg acttatcttc taaataaaat
     3241 tttagtaagt cttgtcctgg catattatac attccatcga tgtagtcttc aacattaaca
     3301 actttaagtc cagcaatttg agttaagggt gttgctctca atgatttcat taatggttca
     3361 atttttaatt tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt gtaatatgca
     3421 cccatttctt taaataaatc ttctaaatag tctactaatg ttttattttg ttttttataa
     3481 aatcaagcag cctctgctat taatatagaa gcttgtattc catctttatc tctagctgag
     3541 tcatcaatta catatccata actttcttca taagcaaaaa caaaatttaa tccgttatct
     3601 tcttctttag caatttctct acccattcat ttaaatccag ttaaagtttt tacaatatta
     3661 actccatatt tttcatgagc gattctatca cccaaatcac ttgttacaaa acttgaatat
     3721 agagccggat tttttggaat gctatttaag cgttttagat ttgataattt tcaatcaatt
     3781 aaaattggtc ctgtttgatt tccatctaat cttacaaaat gaccatcatg ttttattgcc
     3841 attccaaatc tgtcagcatc tgggtcattc ataataataa tatctgcatc atgtttaata
     3901 ccatattcaa gcggtatttt tcatgcagga tcaaattctg gatttggatt tacaacattt
     3961 ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa taacgttata tcctgattca
     4021 cgtaatattt ttggggtaaa tttagttcct gttccattaa ctgcgctaaa aataattttt
     4081 aaatcttttt tagcttcttg ctcttttttg tacgtctctg ttatagtact ctggaaacag
     4141 aatctactat aacaaggcaa aatgccgtgt ttatctcgtc aacttgttgg cgagattttt
     4201 tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     4261 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     4321 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4381 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4441 tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca ccatgaagcg
     4501 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagtgagca agggcgaggc
     4561 agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga
     4621 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4681 gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
     4741 gtacggctcc agggccttca tcaagcaccc cgccgacatc cccgactact ataagcagtc
     4801 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac
     4861 cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg
     4921 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc
     4981 caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg
     5041 cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc
     5101 cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga
     5161 ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat
     5221 ggacgagctg tacaagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga
     5281 tgtcgaagag aatcctggac cgatggccaa gcctttgtct caagaagaat ccaccctcat
     5341 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     5401 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     5461 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     5521 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     5581 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     5641 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     5701 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5761 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5821 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5881 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5941 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6001 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6061 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6121 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6181 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6241 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6301 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6361 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     6421 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6601 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     6661 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     6721 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6781 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     6841 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     6901 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7321 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     7381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     7441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     7561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     7621 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     7681 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     7741 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     7801 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     7861 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     7921 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     7981 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     8041 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     8101 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     8161 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     8221 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     8281 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     8341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8401 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     8461 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8521 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8581 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8641 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8701 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8761 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8821 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     9001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9301 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9541 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9601 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9661 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9721 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9781 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9841 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9901 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9961 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    10021 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10081 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10141 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10201 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10261 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10321 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10381 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10441 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10501 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10561 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10621 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
    10681 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
    10741 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
    10801 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
    10861 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
    10921 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
    10981 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
    11041 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
    11101 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
    11161 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
    11221 attagtcatc gctattacca tgg
//
