LOCUS       sequence-337922-       10200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible overexpression of miRNA or shRNA with dTomato 
            as a reporter fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10200)
  TITLE     Heckl Lab Plasmids
REFERENCE   2  (bases 1 to 10200)
  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"
COMMENT     Sequence Label: SIN40C.TRE.dTomato.miR30n.PGK.sfGFP.P2A.Tet3G
FEATURES             Location/Qualifiers
     source          1..10200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    235..256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(271..276,277..294,295..301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    309..325
                     /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     314..336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     333..349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     333..349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     368..388
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        370..388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        433..812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        869..1060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(892..912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1079..1305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1306..1486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1533..1658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2151..2384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2569..2613
                     /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    2826..2943
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2989..3367
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2997..3015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3033..3051
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3069..3087
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3105..3123
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3141..3159
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3177..3195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3213..3231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3276..3300
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3374..3383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3380..4081
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3513..3531)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3781..3800
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3975..3998
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           4103..4224
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4252..4378
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        4397..4896
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4432..4455)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4829..4848
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      4903..4912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4909..4911,4912..4914,4915..5625)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             5638..5694
                     /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             5695..6441
                     /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"
     misc_feature    6468..7056
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6521..6541)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6939..6950)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7128..7361
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7439..7560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7476..7495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7530..7549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7600..7735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7755..7774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7756..7774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7784..7801)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7784..7800)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7793..7815)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7942..8397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8029..8048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8239..8260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8423..8527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8528..8596,8597..9388)
                     /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(8746..8765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9559..10147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10048..10067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
       61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc
      421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct
      841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
      901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg
     1081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     1141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     2041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     2101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     2161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2761 acccacctcc caaccccgag gggacaattc tagatcttga gacaaatggc agtattcatc
     2821 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2881 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2941 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gtttactccc
     3001 tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag aacgtatgac
     3061 cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta tcagtgatag
     3121 agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca gtttactccc
     3181 tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag aacgtataag
     3241 ctttaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa ccgtcagatc
     3301 gcctggagca attccacaac acttttgtct tataccaact ttccgtacca cttcctaccc
     3361 tcgtaaagga tccgccacca tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg
     3421 cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg
     3481 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     3541 cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt
     3601 gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg
     3661 ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct
     3721 gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg
     3781 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg
     3841 cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta
     3901 cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta
     3961 ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     4021 gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg agctgtacaa
     4081 gtgagcggcc gcatattgct gtttgaatga ggcttcagta ctttacagaa tcgttgcctg
     4141 cacatcttgg aaacacttgc tgggattact tcttcaggtt aacccaacag aaggctaaag
     4201 aaggtatatt gctgttgaca gtgagcggga gacgtgatta ccgtctcttg cctactgcct
     4261 cggacttcaa ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc
     4321 tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt
     4381 tacgcgtaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     4441 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     4501 acatccaccg gaaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     4561 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     4621 caaatggaag tagcacgtgt cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     4681 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     4741 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     4801 ggcgcccgaa ggtccaccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     4861 gcgctgttct cctcttcctc atctccgggc ctttcggcta gcgccaccat gggatctaag
     4921 ggcgaggaac tgttcaccgg cgtggtgccc atcctggtgg aactggatgg cgacgtgaac
     4981 ggccacaagt tctctgtgcg gggagagggc gaaggcgacg ccacaaatgg caagctgacc
     5041 ctgaagttca tctgcaccac cggcaagctg cccgtgcctt ggcctaccct cgtgaccaca
     5101 ctgacctacg gcgtgcagtg cttcagcaga taccccgacc atatgaagcg gcacgacttc
     5161 ttcaagagcg ccatgcccga gggctacgtg caggaacgga ccatcagctt caaggacgac
     5221 ggcacctaca agaccagagc cgaagtgaag ttcgagggcg acaccctcgt gaaccggatc
     5281 gagctgaagg gcatcgattt caaagaggac ggcaacatcc tgggccacaa gctggagtac
     5341 aacttcaaca gccacaatgt gtatatcacc gccgacaagc agaagaacgg catcaaggcc
     5401 aacttcaaga tccggcacaa cgtggaagat ggcagcgtgc agctggccga ccactaccag
     5461 cagaacaccc ccatcggaga tggccccgtg ctgctgcccg acaaccacta cctgagcacc
     5521 cagagcgtgc tgagcaagga ccccaacgag aagcgggacc acatggtgct gctggaattc
     5581 gtgaccgccg ctggcatcac ccacggcatg gacgagctgt ataagggagg aagcggcgcc
     5641 accaatttca gcctgctgaa acaggccggc gacgtggaag agaaccctgg ccctatgagc
     5701 cggctggaca agagcaaagt gatcaacagc gccctggaac tgctgaacgg cgtgggcatc
     5761 gagggcctga ccacccggaa gctggcccag aaactgggcg tggaacagcc caccctgtac
     5821 tggcacgtga agaacaagcg ggccctgctg gacgccctgc ccatcgagat gctggaccgg
     5881 caccacaccc acagctgccc tctggaaggc gagagctggc aggacttcct gcggaacaac
     5941 gccaagagct acagatgcgc cctgctgagc caccgggacg gcgccaaagt gcacctgggc
     6001 accagaccca ccgagaagca gtacgagaca ctggaaaacc agctggcctt cctgtgccag
     6061 cagggcttca gcctggaaaa cgccctgtac gccctgagcg ccgtgggcca cttcaccctg
     6121 ggctgcgtgc tggaagaaca ggaacaccag gtcgccaaag aggaaagaga gacacccacc
     6181 accgacagca tgccccccct gctgaagcag gccatcgagc tgttcgacag acagggcgcc
     6241 gagcccgcct tcctgttcgg cctggaactg atcatctgcg gcctcgagaa gcagctgaag
     6301 tgcgagagcg gcggacccac cgacgccctg gacgacttcg acctggacat gctgcccgcc
     6361 gatgccctgg atgattttga tctggatatg ctccctgctg acgctctcga tgacttcgat
     6421 ctcgatatgc tgccaggcta atgtacaagt gatcgcgtta agtcgacaat caacctctgg
     6481 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     6541 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6601 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6661 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6721 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6781 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6841 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6901 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6961 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     7021 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta
     7081 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     7141 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     7201 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     7261 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     7321 atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg
     7381 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     7441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7561 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     7621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     7681 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     7741 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     7801 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     7861 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     7921 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7981 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8041 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8101 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     8161 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     8221 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8281 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8341 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     8401 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     8461 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     8521 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     8581 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     8641 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     8701 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     8761 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     8821 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     8881 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     8941 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     9001 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     9061 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     9121 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     9181 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     9241 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     9301 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     9361 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     9421 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     9481 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     9541 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     9601 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     9661 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     9721 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     9781 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     9841 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     9901 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     9961 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
    10021 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
    10081 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
    10141 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
//
