LOCUS       sequence-459374-       14606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dox-inducible ancBE4max-NG for mammalian cells. piggyBac 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14606)
  AUTHORS   Osgood NRB, Zawalick NM, Sawyer CB, Cowan QT, Gu S, Mawson SJ, 
            Ranzau BL, Li L, Gymrek M, Goren A, Komor AC
  TITLE     Genome editing with programmable base editors in human cells
  JOURNAL   Methods in Enzymology, February 5, 2025
REFERENCE   2  (bases 1 to 14606)
  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: "Methods in 
            Enzymology, February 5, 2025"
COMMENT     Sequence Label: piggyBac-ancBE4-NG
FEATURES             Location/Qualifiers
     source          1..14606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1744..1766
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1758..1775
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1759..1775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1905..1967
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2416..2794
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2424..2442
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2460..2478
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2496..2514
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2532..2550
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2568..2586
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2604..2622
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2640..2658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2703..2727
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2863..2883
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3664..7764
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7795..8043
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8074..8322
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8365..8385
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8395..8451
                     /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             8452..9159
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8597..8615)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8865..8884
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9172..9192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9201..9218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9241..9258)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9247..9471
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        9732..9943
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             9956..10224
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             10258..11001
                     /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             11002..11055
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             11077..12096
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             11077..12096
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     primer_bind     complement(12153..12173)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    12207..12328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12244..12263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12298..12317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   13091..13125
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(13616..13632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13616..13632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13629..13651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13640..13656
                     /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(13664..13670,13671..13688,13689..13694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13709..13730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13847..13864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14018..14606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14098..14117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1741 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt
     1801 ttgaacccgt ggaggacggg cagactcgcg gtgcaaatgt gttttacagc gtgatggagc
     1861 agatgaagat gctcgacacg ctgcagaaca cgcagctaga ttaaccctag aaagataatc
     1921 atattgtgac gtacgttaaa gataatcatg tgtaaaattg acgcatgtgt tttatcggtc
     1981 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
     2041 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
     2101 caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc
     2161 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     2221 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     2281 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     2341 gagcctgcag acacctgggg ggatacgggg aaaaggcctc cacggccact agtctttcgt
     2401 cttcaagaat tcctcgagtt tactccctat cagtgataga gaacgtatga agagtttact
     2461 ccctatcagt gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat
     2521 aaggagttta ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga
     2581 tagagaacgt atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact
     2641 ccctatcagt gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc
     2701 agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat
     2761 accaactttc cgtaccactt cctaccctcg taaaggtcta gagctagcga attcgaattt
     2821 gccaccatga aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa
     2881 gtcagcagtg aaaccggacc agtggcagtg gacccaaccc tgaggagacg gattgagccc
     2941 catgaatttg aagtgttctt tgacccaagg gagctgagga aggagacatg cctgctgtac
     3001 gagatcaagt ggggcacaag ccacaagatc tggcgccaca gctccaagaa caccacaaag
     3061 cacgtggaag tgaatttcat cgagaagttt acctccgagc ggcacttctg cccctctacc
     3121 agctgttcca tcacatggtt tctgtcttgg agcccttgcg gcgagtgttc caaggccatc
     3181 accgagttcc tgtctcagca ccctaacgtg accctggtca tctacgtggc ccggctgtat
     3241 caccacatgg accagcagaa caggcagggc ctgcgcgatc tggtgaattc tggcgtgacc
     3301 atccagatca tgacagcccc agagtacgac tattgctggc ggaacttcgt gaattatcca
     3361 cctggcaagg aggcacactg gccaagatac ccacccctgt ggatgaagct gtatgcactg
     3421 gagctgcacg caggaatcct gggcctgcct ccatgtctga atatcctgcg gagaaagcag
     3481 ccccagctga catttttcac cattgctctg cagtcttgtc actatcagcg gctgcctcct
     3541 catattctgt gggctacagg cctgaagtct ggaggatcta gcggaggatc ctctggcagc
     3601 gagacaccag gaacaagcga gtcagcaaca ccagagagca gtggcggcag cagcggcggc
     3661 agcgacaaga agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg
     3721 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     3781 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     3841 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     3901 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     3961 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     4021 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     4081 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     4141 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     4201 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     4261 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     4321 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     4381 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     4441 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     4501 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     4561 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     4621 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     4681 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     4741 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     4801 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     4861 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     4921 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     4981 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     5041 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     5101 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     5161 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     5221 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     5281 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     5341 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     5401 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     5461 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     5521 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     5581 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     5641 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     5701 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     5761 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     5821 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     5881 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     5941 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     6001 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     6061 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     6121 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     6181 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     6241 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     6301 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     6361 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     6421 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     6481 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     6541 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     6601 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     6661 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     6721 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     6781 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     6841 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     6901 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     6961 cagacaggcg gcttcagcaa agagtctatc cgtcccaaga ggaacagcga taagctgatc
     7021 gccagaaaga aggactggga ccctaagaag tacggcggct tcgtgagccc caccgtggcc
     7081 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     7141 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     7201 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     7261 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc ccggttcctg
     7321 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     7381 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     7441 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     7501 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     7561 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     7621 cctagggcct tcaagtactt tgacaccacc atcgaccgga aggtgtaccg tagcaccaaa
     7681 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     7741 gacctgtctc agctgggagg tgacagcggc gggagcggcg ggagcggggg gagcactaat
     7801 ctgagcgaca tcattgagaa ggagactggg aaacagctgg tcattcagga gtccatcctg
     7861 atgctgcctg aggaggtgga ggaagtgatc ggcaacaagc cagagtctga catcctggtg
     7921 cacaccgcct acgacgagtc cacagatgag aatgtgatgc tgctgacctc tgacgccccc
     7981 gagtataagc cttgggccct ggtcatccag gattctaacg gcgagaataa gatcaagatg
     8041 ctgagcggag gatccggagg atctggaggc agcaccaacc tgtctgacat catcgagaag
     8101 gagacaggca agcagctggt catccaggag agcatcctga tgctgcccga agaagtcgaa
     8161 gaagtgatcg gaaacaagcc tgagagcgat atcctggtcc ataccgccta cgacgagagt
     8221 accgacgaaa atgtgatgct gctgacatcc gacgccccag agtataagcc ctgggctctg
     8281 gtcatccagg attccaacgg agagaacaaa atcaaaatgc tgtctggcgg ctcaaaaaga
     8341 accgccgacg gcagcgaatt cgagcccaag aagaagagga aagtcggaag cggagctact
     8401 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatggtgagc
     8461 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     8521 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     8581 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     8641 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     8701 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     8761 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     8821 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     8881 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     8941 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     9001 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     9061 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     9121 ggccgccact ccaccggcgg catggacgag ctgtacaagt ctggtggttc tcccaagaag
     9181 aagaggaaag tctaaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     9241 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     9301 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     9361 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     9421 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     9481 cggaaagaac cagctggggc tataaacgcg gggttcggtc ccagggctgg cactctgtcg
     9541 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     9601 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     9661 ccatagcaga tctgcgaaat cggatccagc taaatctaag tggatctgcg atcgctccgg
     9721 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     9781 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     9841 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     9901 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     9961 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
    10021 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
    10081 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
    10141 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
    10201 ttctgttctg cgccgttaca gatctaagct gtgaccggcg cctactctag agccgccatg
    10261 tctagactgg acaagagcaa agtcataaac tctgctctgg aattactcaa tggagtcggt
    10321 atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg
    10381 tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac
    10441 aggcatcata cccactcctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac
    10501 aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc
    10561 ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt
    10621 cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca
    10681 ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct
    10741 accaccgatt ctatgccccc acttctgaaa caagcaattg agctgttcga ccggcaggga
    10801 gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta
    10861 aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca
    10921 gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt
    10981 gaccttgaca tgctccccgg ggagggcaga ggaagtcttc taacatgcgg tgacgtggag
    11041 gagaatcccg gccctgaatt catggataga tccggaaagc ctgaactcac cgcgacgtct
    11101 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
    11161 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
    11221 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
    11281 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
    11341 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
    11401 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
    11461 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
    11521 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
    11581 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
    11641 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
    11701 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
    11761 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
    11821 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
    11881 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
    11941 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
    12001 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
    12061 cgacgcccca gcactcgtcc gagggcaaag gaataagcca aggaacctct ggattacaaa
    12121 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
    12181 gctgctttaa tgcctttgta tcagttaact tgtttattgc agcttataat ggttacaaat
    12241 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    12301 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat
    12361 tagtctatca gaagctcatc tggtctccct tccgggggac aagacatccc tgtttaatat
    12421 ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc
    12481 agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc
    12541 cccggaattg actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc
    12601 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc
    12661 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg
    12721 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa
    12781 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat
    12841 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
    12901 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
    12961 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
    13021 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
    13081 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat
    13141 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg
    13201 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag
    13261 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg
    13321 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg
    13381 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc
    13441 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac
    13501 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag
    13561 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat
    13621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    13681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    13741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    13801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    13861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    13921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    13981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgctcccctg
    14041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    14101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    14161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    14221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    14281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    14341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    14401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    14461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    14521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    14581 ttacgcgcag aaaaaaagga tctcaa
//
