LOCUS       sequence-376601-        9382 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector to co-express a mouse Itgb1 spsgRNA (sg2-Itgb1) 
            with NLS-mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9382)
  AUTHORS   Sekiguchi R, Mehlferber MM, Matsumoto K, Wang S
  TITLE     Efficient Gene Knockout in Salivary Gland Epithelial Explant 
            Cultures.
  JOURNAL   J Dent Res. 2023 Feb;102(2):197-206. doi: 10.1177/00220345221128201.
            Epub 2022 Nov 10.
  PUBMED    36366748
REFERENCE   2  (bases 1 to 9382)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1177/00220345221128201"; journalName: "J Dent Res"; date: 
            "2023-02"; volume: "102"; issue: "2"; pages: "197-206"
COMMENT     Sequence Label: 
            pW402-lenti-sg2-mmItgb1-pEF1s-NLS-mScarlet-I-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..9382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..201
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     156..176
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             219..399
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    446..571
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1064..1297
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1482..1526
                     /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    1824..1941
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1992..2240
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1992..2012
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2163..2182
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2261..2336
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2398..2609
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2637..2684
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2685..3377
                     /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(2815..2833)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3083..3102
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3387..3443
                     /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             3444..3842
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3858..4446
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3911..3931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4329..4340)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4518..4751
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4777..4794)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4783..5007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5053..5481
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5140..5159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5350..5371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5490..5510)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5495..5824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5675..5810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5737..5756
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5872..5919
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5938..6312
                     /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    6442..6563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6479..6498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6533..6552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6612..6628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6612..6628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6625..6647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6636..6652
                     /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(6660..6666,6667..6684,6685..6690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6705..6726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6843..6860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7014..7602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7094..7113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7773..8564,8565..8633))
                     /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     8396..8415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8634..8738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8813..8832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9004..9382,1)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
       61 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      121 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      181 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
      241 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      301 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      361 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      421 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      481 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      541 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      601 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      661 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
      721 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
      781 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
      841 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
      901 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
      961 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1021 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1081 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1141 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1201 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1261 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1321 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1381 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     1441 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     1501 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     1561 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     1621 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     1681 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     1741 acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga
     1801 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     1861 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     1921 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta
     1981 attaaggtac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     2041 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     2101 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     2161 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     2221 tgtggaaagg acgaaacacc gcggagaatg tatacaagca gttttagagc tagaaatagc
     2281 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2341 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2401 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2461 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2521 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2581 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgaagc
     2641 gacctgccgc cacaaagaag gctggacagg ctaagaagaa gaaagtgagc aagggcgagg
     2701 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
     2761 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2821 tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     2881 tgtacggctc cagggccttc atcaagcacc ccgccgacat ccccgactac tataagcagt
     2941 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga
     3001 ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg
     3061 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt
     3121 ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc
     3181 gcctgaagga cggcggccgc tacctggcgg acttcaagac cacctacaag gccaagaagc
     3241 ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg
     3301 aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca
     3361 tggacgagct gtacaaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag
     3421 atgtcgaaga gaatcctgga ccgatggcca agcctttgtc tcaagaagaa tccaccctca
     3481 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     3541 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     3601 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     3661 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     3721 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     3781 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     3841 aaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3901 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3961 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4021 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4081 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4141 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4201 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4261 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4321 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4381 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4441 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     4501 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     4561 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4621 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4681 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4741 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     4801 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4861 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4921 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4981 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     5041 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5101 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5161 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5221 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5281 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5341 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5401 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5461 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5521 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5581 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5641 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5701 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5761 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5821 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     5881 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     5941 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     6001 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     6061 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     6121 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     6181 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     6241 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     6301 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6361 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6421 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     6481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6541 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     6601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8761 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     8821 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8881 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8941 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     9001 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     9061 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     9121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     9181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     9241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     9301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     9361 tattagtcat cgctattacc at
//
