LOCUS       sequence-322808-       12623 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For making a cell line that expresses doxycyline-inducible RNAi 
            suppressor protein B2 for increasing titer of lentivirus with 
            bidirectional promoters during virus production..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12623)
  AUTHORS   Choi SH, Reeves RE, Romano Ibarra GS, Lynch TJ, Shahin WS, Feng Z, 
            Gasser GN, Winter MC, Evans TIA, Liu X, Luo M, Zhang Y, Stoltz DA, 
            Devor EJ, Yan Z, Engelhardt JF
  TITLE     Detargeting Lentiviral-Mediated CFTR Expression in Airway Basal 
            Cells Using miR-106b.
  JOURNAL   Genes (Basel). 2020 Oct 6;11(10). pii: genes11101169. doi: 
            10.3390/genes11101169.
  PUBMED    33036232
REFERENCE   2  (bases 1 to 12623)
  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"; journalName: "Genes 
            (Basel)."; date: "2020-10-6"; volume: "11(10). pii: genes11101169. 
            doi"; pages: " 10.3390/genes11101169"
COMMENT     Sequence Label: pTripZ-B2
FEATURES             Location/Qualifiers
     source          1..12623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3373..4584
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4421..4440
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(4597..4630)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             4637..5344
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(5345..5378)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    5401..5973
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5568..5585)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5795..5814
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5976..6578
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6475..6495
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6594..7182
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6647..6667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7065..7076)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7389..7622
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7723..7740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7729..7953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7999..8427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8086..8105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8296..8317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8436..8456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8441..8770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8621..8756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8683..8702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9978..10099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10148..10164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10148..10164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10161..10183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10172..10188
                     /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(10196..10202,10203..10220,10221..10226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10241..10262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10379..10396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10550..11138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10630..10649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11309..12100,12101..12169))
                     /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     11932..11951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12170..12274)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12322..12456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggtgcc
     3001 gccaccatgc caagcaaact cgcgctaatc caggaacttc ccgaccgcat tcaaacggcg
     3061 gtggaagcag ccatgggaat gagctaccaa gacgcaccga acaacgtgcg cagggacctc
     3121 gacaacctgc acgcttgcct aaacaaggca aaactaacgg taagtcggat ggtaacatca
     3181 ctgctggaga aacccagcgt ggtggcatac ctagagggaa aggcccccga ggaggcaaaa
     3241 ccaacactcg aagaacgcct ccgaaagctg gagctcagcc acagccttcc aacaaccgga
     3301 agtgaccccc cacccgcaaa actgtaggtg gctcttagga gcacccacac ccgttctagc
     3361 ccgaaaacgc gtggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca
     3421 cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg tcctgatcct tccgcccgga
     3481 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3541 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3601 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3661 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3721 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3781 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3841 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     3901 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     3961 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     4021 tcttgaggcc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca
     4081 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     4141 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     4201 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     4261 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     4321 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     4381 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     4441 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     4501 tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg
     4561 ccgtttttgg cttttttgtt agacgctagc ggatccataa cttcgtatag tatacattat
     4621 acgaagttat gccaccatgt ctaggctgga caagagcaaa gtcataaacg gagctctgga
     4681 attactcaat ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg
     4741 agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct
     4801 gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg
     4861 gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga
     4921 cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa
     4981 tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc
     5041 cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa
     5101 agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga
     5161 gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg
     5221 tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt
     5281 tgacttagac atgctcccag ccgatgccct tgacgatttt gaccttgaca tgctccccgg
     5341 gtaaataact tcgtatagta tacattatac gaagttatgg atccgcggcc gcaaattccg
     5401 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     5461 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     5521 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     5581 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     5641 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     5701 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     5761 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5821 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     5881 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     5941 ggacgtggtt ttcctttgaa aaacacgata ataccatggc caccgagtac aagcccacgg
     6001 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     6061 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     6121 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     6181 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     6241 tcgccgagat cggctcgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     6301 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     6361 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     6421 tggaggcggc cgagcgcgct ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     6481 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6541 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt tcgcgtctgg aacaatcaac
     6601 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6661 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6721 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6781 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6841 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6901 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6961 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     7021 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     7081 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     7141 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     7201 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     7261 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     7321 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     7381 gaggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     7441 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     7501 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     7561 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     7621 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     7681 agagagtgag aggccttgac attgtttaaa cccgctgatc agcctcgact gtgccttcta
     7741 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     7801 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     7861 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     7921 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     7981 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     8041 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     8101 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     8161 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     8221 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     8281 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     8341 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     8401 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     8461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     8521 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     8581 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     8641 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     8701 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     8761 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat
     8821 gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     8881 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     8941 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     9001 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     9061 ggaattaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     9121 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     9181 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     9241 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     9301 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     9361 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     9421 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     9481 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     9541 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     9601 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     9661 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     9721 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     9781 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     9841 aggaatagca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     9901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     9961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
    10021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
    10081 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
    10141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
    10201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
    10261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
    10321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
    10381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
    10441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
    10501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
    10561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
    10621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
    10681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
    10741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
    10801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
    10861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
    10921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
    10981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
    11041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    11101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    11161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    11221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    11281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    11341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    11401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    11461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    11521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    11581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    11641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    11701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    11761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    11821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    11881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    11941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    12001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    12061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    12121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    12181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    12241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
    12301 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
    12361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    12421 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    12481 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    12541 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    12601 atatgtacta caaacttagt agt
//
