LOCUS       sequence-386853-       12589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible 3xFLAG-tagged TWIST2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12589)
  AUTHORS   Dutta AB, Lank DS, Przanowska RK, Przanowski P, Wang L, Nguyen B, 
            Walavalkar NM, Duarte FM, Guertin MJ
  TITLE     Kinetic networks identify TWIST2 as a key regulatory node in 
            adipogenesis.
  JOURNAL   Genome Res. 2023 Feb 21;33(3):314-31. doi: 10.1101/gr.277559.122.
  PUBMED    36810156
REFERENCE   2  (bases 1 to 12589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1101/gr.277559";
            journalName: "Genome Res"; date: "2023-02-21- 21"; volume: "33"; 
            issue: "3"; pages: "314-31"
COMMENT     Sequence Label: pSLIK 3xFLAG-TWIST2 neo
FEATURES             Location/Qualifiers
     source          1..12589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(124..143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        315..694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        696..894
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             912..1092
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1139..1264
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1757..1990
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2175..2219
                     /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    2517..2634
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2691..2715
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2747..3061
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2755..2773
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2791..2809
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2826..2844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2862..2880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2898..2916
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2933..2951
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2969..2987
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3034..3058
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3070..3135
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(3660..3684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3734..4945
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4782..4801
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5004..5621,5622..5627,5628..5663,5664..5666,5667..5702,
                     5703..5705,5706..5741,5742..5750)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    5803..6376
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5970..5987)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6197..6216
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6392..7186
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6392..7186
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6446..6465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7056..7075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    7354..7942
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7407..7427)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7825..7836)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7945..7961)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7946..7962)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8471..8651
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8677..8694)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8683..8907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8953..9381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9040..9059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9250..9271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9390..9410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9395..9724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9575..9710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9637..9656
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9772..9819
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9838..10212
                     /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    10342..10463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10379..10398)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10433..10452
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10512..10528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10512..10528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10525..10547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10536..10552
                     /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(10560..10566,10567..10584,10585..10590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10605..10626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10743..10760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10914..11502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10994..11013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11673..12464,12465..12533))
                     /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     12296..12315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(12534..12589,1..49))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
       61 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
      121 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
      181 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
      241 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      301 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      361 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      421 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      481 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      541 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      601 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      661 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      721 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      781 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      841 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
      901 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      961 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     1021 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1081 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     1141 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     1201 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     1261 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     1321 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     1381 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     1441 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     1501 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     1561 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     1621 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1681 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1741 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1801 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1861 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1921 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1981 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     2041 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     2101 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     2161 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     2221 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     2281 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2341 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2401 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2461 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     2521 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2581 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2641 tattacaggg acagcagaga tccagtttgg ttaattctag acagttatca acaagtttgt
     2701 acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg gatcccgagt ttactcccta
     2761 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgatgtcgag
     2821 tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga
     2881 acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt tatccctatc
     2941 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagg
     3001 taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3061 cactagatgg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     3121 gatgacgatg acaagactag tatggaggag ggctccagct cgcccgtgtc ccccgtggac
     3181 agcctgggca ccagcgagga ggagctcgag aggcagccca agcgcttcgg ccggaagcgg
     3241 cgctacagca agaagtcgag cgaagatggc agcccgaccc cgggcaagcg cggcaagaag
     3301 ggcagcccca gcgcgcagtc cttcgaggag ctgcagagcc agcgcatcct ggccaacgtg
     3361 cgcgagcgcc agcgcaccca gtcgctcaac gaggccttcg cggcgctgcg caagatcatc
     3421 cccacgctgc cctctgacaa gctgagcaag atccagacgc tcaagctggc cgccaggtac
     3481 atagacttcc tctaccaggt cctgcagagc gacgagatgg acaataagat gaccagctgc
     3541 agctacgtgg cccacgagcg cctcagctac gccttctccg tgtggcgcat ggagggcgcg
     3601 tggtccatgt ccgcctccca ctagcaattg gaagactaat gcgttttcga gatatctaga
     3661 cccagctttc ttgtacaaag tggttgataa catctcgagc cataaagatg gttaattaac
     3721 ccacccaaga tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc
     3781 acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg
     3841 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     3901 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     3961 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     4021 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     4081 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     4141 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     4201 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     4261 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     4321 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     4381 gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc
     4441 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     4501 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     4561 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     4621 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     4681 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     4741 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     4801 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     4861 atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg
     4921 gccgtttttg gcttttttgt tagacgaagc ttggtaccga gctcggatct ccaccccgta
     4981 ccggtcctgc agtcgaattc accatgtcta gactggacaa gagcaaagtc ataaacggag
     5041 ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa
     5101 agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg
     5161 atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc ctggaaggcg
     5221 agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac
     5281 atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc
     5341 tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg
     5401 ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag
     5461 tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt ctgagacaag
     5521 caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa
     5581 tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg
     5641 acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc
     5701 tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa ctaagtaagg
     5761 atccgcggcc gcactagagg aattccgccc ctctcccccc cccccctctc cctccccccc
     5821 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     5881 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     5941 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     6001 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     6061 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     6121 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     6181 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     6241 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     6301 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     6361 aaaaacacga tgataagctt accggtccac catgattgaa caagatggat tgcacgcagg
     6421 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     6481 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     6541 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     6601 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6661 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6721 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6781 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6841 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6901 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6961 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     7021 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     7081 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     7141 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgatgta caagtaaagc
     7201 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     7261 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgttt
     7321 agtccctccc aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     7381 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     7441 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     7501 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     7561 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     7621 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     7681 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     7741 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     7801 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     7861 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     7921 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     7981 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     8041 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     8101 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     8161 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     8221 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     8281 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     8341 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     8401 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     8461 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     8521 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     8581 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     8641 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     8701 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     8761 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     8821 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     8881 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     8941 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     9001 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     9061 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     9121 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     9181 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     9241 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     9301 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     9361 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     9421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     9481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     9541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     9601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     9661 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     9721 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     9781 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     9841 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     9901 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     9961 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
    10021 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
    10081 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
    10141 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
    10201 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
    10261 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
    10321 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
    10381 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10441 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
    10501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    10561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    10621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    10681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    10741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    10801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    10861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    10921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    10981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    11041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    11101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    11161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    11221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    11281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    11341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    11401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    11461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    11521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    11581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    11641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    11701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    11761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    11821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    11881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    11941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    12001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    12061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    12121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    12181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    12241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    12301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    12361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    12421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    12481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    12541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcg
//
