LOCUS       sequence-492552-       10268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZFa Lentivirus expressing VP16-ZF43-2A-TagBFP from the TET promoter 
            with rtTA expressed divergently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10268)
  AUTHORS   Kabaria SR, Bae Y, Ehmann ME, Lende-Dorn BA, Beitz AM, Peterman EL, 
            Love KS, Ploessl DS, Galloway KE
  TITLE     Programmable promoter editing for precise control of transgene 
            expression.
  JOURNAL   Nat Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y.
  PUBMED    41083901
REFERENCE   2  (bases 1 to 10268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y."
COMMENT     Sequence Label: LentiX1-TET-VP16-ZF43-2A-tagBFP-bGH_DIVrtTA_pA...
FEATURES             Location/Qualifiers
     source          1..10268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..50
                     /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    317..433
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    460..684
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(696..1397)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             complement(1410..1466)
                     /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             complement(2019..2039)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2046..2111)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      2111..2120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2176..2554)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2312..2330
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2348..2366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2384..2402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2420..2438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2456..2474
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2492..2510
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2528..2546
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2735..2754)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        2864..3075
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3088..3831
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             3841..3897
                     /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             3898..4608
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    4622..5210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4675..4695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5093..5104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5302..5535
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5607..5728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5644..5663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5698..5717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5768..5903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5923..5942)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5924..5942)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5952..5969)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5952..5968)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5961..5983)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6110..6565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6197..6216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6407..6428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6591..6695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6696..6764,6765..7556)
                     /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(6914..6933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7727..8315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8216..8235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8469..8486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8603..8624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8639..8644,8645..8662,8663..8669)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8677..8693
                     /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     8682..8704
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8701..8717
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8701..8717
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8736..8756
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8738..8756
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8784..9010
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9011..9191
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9238..9363
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9856..10089
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
       61 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
      121 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
      181 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
      241 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc
      301 gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
      361 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
      421 aattcaaaat tttatcgtcc agtgtggtgg ctactagcgc catagagccc accgcatccc
      481 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
      541 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
      601 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
      661 atggctggca actagaaggc acaggtgcaa cgttgttaat taagcttgtg ccccagtttg
      721 ctagggaggt cgcagtatct ggccactgcc acctcgtgct gctcgacgta cgtttcgttg
      781 ttggcctcct tgattctttc cagtctgtag tccacatagt agacgccagg catcttgagg
      841 ttcttagcgg gtttcttgga tctatatgtg gtcttgatgt ttgcgatcag atggctcccg
      901 cccacgagct tcagggccat gtcgtttctg ccttccaggc cgccgtcagc ggggtacagc
      961 gtctcggtga aggcctccca gccgagtgtt ttcttctgca tcacagggcc gttggatgtg
     1021 aagttcaccc ctctgatctt gacgttgtag atgaggcagc cgtcctggag gctggtgtcc
     1081 tgggtagcgg tcagcacgcc cccgtcttcg tatgtggtga ctctctccca tgtgaagccc
     1141 tcagggaagg actgcttgaa gaagtcgggg atgccctggg tgtggttgat gaaggtcttg
     1201 ctgccgtaga ggaagctagt agccaggatg tcgaaggcga aggggagagg gccgccctcg
     1261 accaccttga ttctcatggt ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg
     1321 cacttgaagt gatggttgtc cacggtgccc tccatgtaca gcttcatgtg catgttctcc
     1381 ttaatcagct cgctcatact tcctggtgca ggtccagggt tctcctccac gtctccagcc
     1441 tgcttcagca ggctgaagtt agtagctccg cttccagatc ctctcaggtg ggtttttagg
     1501 tgccggttca ggttgtcgcg gcgactgaag ttgcgcatgc atattcggca ttggaatggc
     1561 ttctcgccgg tgtgcgtacg tagatgcccc gccaagtgct ccttctggga gaaatttcgc
     1621 atacagatcc gacactgaaa cggtttttca ccggtatgag tacgggtatg cctgtcaagc
     1681 ctgtcctggc gcgaaaagtt ccgcatgcaa atgcgacact ggaaggggcg ctccccgggt
     1741 ctagctccac caccgtattc gtctataccc aaggcatcag taaacatctg ttcaaattcg
     1801 aaatcggcca tgtccaaagc accgtatgga gctgaatcgt gtggtgtgaa accaggtcct
     1861 ggtgaatctc catctcctaa catatccaag tcaaaatcgt ctaaggcatc tgcatgtgcc
     1921 atagctacgt cctcaccgtc caaatgtaac tcatcaccta aagaaacatc ggttggtggg
     1981 gcctgcagtc caccgtcgac gggaacgcca tggataccga ctttgcgctt tttcttagga
     2041 gccattttat cgtcatcgtc tttgtagtca atatcgtggt ccttgtagtc gccatcgtga
     2101 tccttataat ccatggtggc acattacagc gtggatggcg tctccaggcg atctgacggt
     2161 tcactaaacg agctctttac gagggtagga agtggtacgg aaagttggta taagacaaaa
     2221 gtgttgtgga attgctccag gcgatctgac ggttcactaa acgagctctg cttttatagg
     2281 cgcccaccgt acacgcctaa agcttatacg ttctctatca ctgataggga gtaaactgga
     2341 tatacgttct ctatcactga tagggagtaa actgtagata cgttctctat cactgatagg
     2401 gagtaaactg gtcatacgtt ctctatcact gatagggagt aaactcctta tacgttctct
     2461 atcactgata gggagtaaag tctgcatacg ttctctatca ctgataggga gtaaactctt
     2521 catacgttct ctatcactga tagggagtaa actcgaggaa tagtacgaac atggtagcta
     2581 gtggatcctt cgtcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc
     2641 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
     2701 ttcttccccg ataacggaga ccggcacact ggcgcatatc ggtggtcatc atgcgccagc
     2761 tttcatcccc gatatgcacc accgggtaaa gttcacgcat atgaagctgg acatcacctc
     2821 ccacaacctc gaggtggaac agtacgctga tatctcttat actgggcaga gcgcacatcg
     2881 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     2941 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     3001 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3061 gccgccagaa cacagtaatg tgccaccatg agccggctgg acaagagcaa agtgatcaac
     3121 agcgccctgg aactgctgaa cggcgtgggc atcgagggcc tgaccacccg gaagctggcc
     3181 cagaaactgg gcgtggaaca gcccaccctg tactggcacg tgaagaacaa gcgggccctg
     3241 ctggacgccc tgcccatcga gatgctggac cggcaccaca cccacagctg ccctctggaa
     3301 ggcgagagct ggcaggactt cctgcggaac aacgccaaga gctacagatg cgccctgctg
     3361 agccaccggg acggcgccaa agtgcacctg ggcaccagac ccaccgagaa gcagtacgag
     3421 acactggaaa accagctggc cttcctgtgc cagcagggct tcagcctgga aaacgccctg
     3481 tacgccctga gcgccgtggg ccacttcacc ctgggctgcg tgctggaaga acaggaacac
     3541 caggtcgcca aagaggaaag agagacaccc accaccgaca gcatgccccc cctgctgaag
     3601 caggccatcg agctgttcga cagacagggc gccgagcccg ccttcctgtt cggcctggaa
     3661 ctgatcatct gcggcctcga gaagcagctg aagtgcgaga gcggcggacc caccgacgcc
     3721 ctggacgact tcgacctgga catgctgccc gccgatgccc tggatgattt tgatctggat
     3781 atgctccctg ctgacgctct cgatgacttc gatctcgata tgctgccagg cggaagcgga
     3841 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg
     3901 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
     3961 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaatcc gtacatggga
     4021 actcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
     4081 cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccgaa aggcattcct
     4141 gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa
     4201 gatggtggag tcgtcaccgt catgcaggac accagccttg aggatggctg tctcgtttac
     4261 cacgtccaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     4321 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gaggggatac
     4381 actcatatgg cactgaaagt tgatggtggt ggccatctgt cttgctcttt cgtaacaact
     4441 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtatccatgc cgttgatcac
     4501 cgcctggaaa ggttagagga aagtgacaat gaaatgttcg tagtacaacg cgaacacgca
     4561 gttgccaagt tcgccgggct tggtggtggg atggacgagc tgtacaagta acaacgccga
     4621 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4681 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4741 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4801 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4861 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4921 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4981 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5041 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5101 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5161 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc cgctcgctga
     5221 tatagggccc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5281 ttttaaaaga aaagggggga ctggaagggc tagctcactc ccaacgaaga caagatctgc
     5341 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5401 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5461 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5521 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     5581 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     5641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5701 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     5761 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5821 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5881 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     5941 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6001 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6061 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     6121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6241 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6541 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6661 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6721 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6781 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6841 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6901 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6961 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7021 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7081 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7141 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7201 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7261 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7321 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7381 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7441 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7501 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7561 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8401 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8461 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8521 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     8581 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     8641 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     8701 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     8761 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
     8821 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
     8881 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     8941 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     9001 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     9061 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     9121 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     9181 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     9241 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     9301 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     9361 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     9421 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     9481 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     9541 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     9601 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     9661 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     9721 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     9781 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     9841 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     9901 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     9961 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
    10021 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
    10081 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
    10141 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
    10201 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
    10261 aaaccagc
//
