LOCUS       sequence-261470-       11900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 donor vector for inducible gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11900)
  AUTHORS   Qian K, Huang CL, Chen H, Blackbourn LW 4th, Chen Y, Cao J, Yao L, 
            Sauvey C, Du Z, Zhang SC
  TITLE     A simple and efficient system for regulating gene expression in 
            human pluripotent stem cells and derivatives.
  JOURNAL   Stem Cells. 2014 Feb 4. doi: 10.1002/stem.1653.
  PUBMED    24497442
REFERENCE   2  (bases 1 to 11900)
  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: "Stem Cells.
            2014 Feb 4. doi: 10.1002/stem.1653."
COMMENT     Sequence Label: AAVS1-TRE3G-EGFP
FEATURES             Location/Qualifiers
     source          1..11900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    37..840
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    847..872
                     /label=SA
                     /note="splice acceptor site"
     CDS             896..949
                     /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             959..1558
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(959..978)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1455..1475
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1590..1607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1596..1820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1858..1979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1869..1888)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1923..1942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2404..2424)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          2554..2619
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(join(2780..3493,3494..3496,3497..3499))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2783..3493,3494..3496,3497..3499))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2825..2846)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3174..3193
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3433..3454
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3506..3884)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     primer_bind     complement(3573..3597)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3642..3660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3678..3696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3714..3732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3750..3768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3786..3804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3822..3840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3858..3876
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        3968..4271
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4273..4548
                     /label=chicken beta-actin promoter
     intron          4549..5565
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5489..5511
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5573..5592
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5630..6376
                     /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"
     primer_bind     complement(6467..6486)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6532..6587
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6586..6605)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6948..6964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6972..6988
                     /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(6996..7002,7003..7020,7021..7026))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7041..7062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7136..7972
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(8006..8025)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8007..8025)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8032..8049)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8032..8048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8041..8063)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8186..8488
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8367..8386
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8891..8910)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9505..9524
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9885..10676,10677..10745))
                     /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     10508..10527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10869..11457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11358..11377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11611..11628
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11745..11766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11781..11786,11787..11804,11805..11811)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11819..11835
                     /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     11843..11859
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11880..11898
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagctagtc ctgcaggttt aaacgaattc gccctttgct ttctctgacc agcattctct
       61 cccctgggcc tgtgccgctt tctgtctgca gcttgtggcc tgggtcacct ctacggctgg
      121 cccagatcct tccctgccgc ctccttcagg ttccgtcttc ctccactccc tcttcccctt
      181 gctctctgct gtgttgctgc ccaaggatgc tctttccgga gcacttcctt ctcggcgctg
      241 caccacgtga tgtcctctga gcggatcctc cccgtgtctg ggtcctctcc gggcatctct
      301 cctccctcac ccaaccccat gccgtcttca ctcgctgggt tcccttttcc ttctccttct
      361 ggggcctgtg ccatctctcg tttcttagga tggccttctc cgacggatgt ctcccttgcg
      421 tcccgcctcc ccttcttgta ggcctgcatc atcaccgttt ttctggacaa ccccaaagta
      481 ccccgtctcc ctggctttag ccacctctcc atcctcttgc tttctttgcc tggacacccc
      541 gttctcctgt ggattcgggt cacctctcac tcctttcatt tgggcagctc ccctaccccc
      601 cttacctctc tagtctgtgc tagctcttcc agccccctgt catggcatct tccaggggtc
      661 cgagagctca gctagtcttc ttcctccaac ccgggcccct atgtccactt caggacagca
      721 tgtttgctgc ctccagggat cctgtgtccc cgagctggga ccaccttata ttcccagggc
      781 cggttaatgt ggctctggtt ctgggtactt ttatctgtcc cctccacccc acagtggggc
      841 aagcttctga cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg
      901 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta ggctcgagat
      961 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     1021 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     1081 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     1141 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     1201 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     1261 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     1321 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     1381 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     1441 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     1501 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatc
     1561 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1621 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1681 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1741 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1801 ggatgcggtg ggctctatgg gtctcgacat tgattattga ctaggccgca gacatgataa
     1861 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1921 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1981 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2041 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctctag tcaaggcact
     2101 atacatcaaa tattccttat taaccccttt acaaattaaa aagctaaagg tacacaattt
     2161 ttgagcatag ttattaatag cagacactct atgcctgtgt ggagtaagaa aaaacagtat
     2221 gttatgatta taactgttat gcctacttat aaaggttaca gaatattttt ccataatttt
     2281 cttgtatagc agtgcagctt tttcctttgt ggtgtaaata gcaaagcaag caagagttct
     2341 attactaaac acagcatgac tcaaaaaact tagcaattct gaaggaaagt ccttggggtc
     2401 ttctaccttt ctcttctttt ttggaggagt agaatgttga gagtcagcag tagcctcatc
     2461 atcactagat ggcatttctt ctgagcaaaa caggttttcc tcattaaagg cattccacca
     2521 ctgctcccat tcatcagttc cataggttgg aatctaaaat acacaaacaa ttagaatcag
     2581 tagtttaaca cattatacac ttaaaaattt tatatttacc ttagagcttt aaatctctgt
     2641 aggtagtttg tccaattatg tcacaccaca gaagtaaggt tccttcacaa agatcctcta
     2701 gagtattaca atagctaaga atttcgtcat cgctgaatac agttacattg gatccctgca
     2761 ggctagccat atgacgcgtt tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     2821 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     2881 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     2941 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     3001 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     3061 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     3121 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     3181 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     3241 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     3301 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     3361 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     3421 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     3481 cctcgccctt gctcaccatg tcgactttac gagggtagga agtggtacgg aaagttggta
     3541 taagacaaaa gtgttgtgga attgctccag gcgatctgac ggttcactaa acgagctctg
     3601 cttttatagg cgcccaccgt acacgcctaa agcttatacg ttctctatca ctgataggga
     3661 gtaaactgga tatacgttct ctatcactga tagggagtaa actgtagata cgttctctat
     3721 cactgatagg gagtaaactg gtcatacgtt ctctatcact gatagggagt aaactcctta
     3781 tacgttctct atcactgata gggagtaaag tctgcatacg ttctctatca ctgataggga
     3841 gtaaactctt catacgttct ctatcactga tagggagtaa actcgaggaa ttcttgaaga
     3901 cgaaaactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     3961 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4021 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4081 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4141 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4201 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4261 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     4321 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     4381 gggggggggg ggsgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     4441 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     4501 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     4561 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
     4621 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
     4681 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     4741 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     4801 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     4861 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     4921 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     4981 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     5041 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     5101 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     5161 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     5221 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     5281 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     5341 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     5401 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     5461 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     5521 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     5581 ctggttattg tgctgtctca tcattttggc aaagaattcg tcgagcacca tgtctagact
     5641 ggacaagagc aaagtcataa actctgctct ggaattactc aatggagtcg gtatcgaagg
     5701 cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca
     5761 cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca
     5821 tacccactcc tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa
     5881 gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg
     5941 cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg
     6001 cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg
     6061 cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga
     6121 ttctatgccc ccacttctga aacaagcaat tgagctgttc gaccggcagg gagccgaacc
     6181 tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga
     6241 aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc
     6301 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga
     6361 catgctcccc gggtaaggcg cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg
     6421 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     6481 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     6541 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     6601 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     6661 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     6721 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     6781 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     6841 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     6901 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     6961 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7021 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7081 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcg acagtactaa gctttactag
     7141 ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta gtctcctgat
     7201 attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca cacccccatt
     7261 tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat ggagccagag
     7321 aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat gcgtgacctg
     7381 cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg ctctgacgcg
     7441 gctgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc ccaagaggag
     7501 aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact ttggctcttc
     7561 acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct gtgagataag
     7621 gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa
     7681 actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg cctctactcc
     7741 ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg ggacatattc
     7801 ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct gggtttgagt
     7861 ccttggcaag cccaggagag gcgctcaggc ttccctgtcc cccttcctcg tccaccatct
     7921 catgcccctg gctctcctgc cccttcccta caggggttcc tggctctgct ctaagggcga
     7981 attcgcggcc gctaaattca attcgcccta tagtgagtcg tattacaatt cactggccgt
     8041 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     8101 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     8161 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
     8221 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
     8281 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     8341 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     8401 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     8461 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt
     8521 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     8581 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     8641 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     8701 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     8761 ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     8821 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     8881 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     8941 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     9001 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     9061 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     9121 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     9181 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     9241 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     9301 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     9361 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat gcatggcgat gcctgcttgc
     9421 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     9481 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     9541 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     9601 tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac aatttcctga
     9661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
     9721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     9781 ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    10021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    10081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    10141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    10201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    10261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    10321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    10381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac
    10441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    10501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    10561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    10621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    10681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    10741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgacc
    10801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    10861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    10921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    10981 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
    11041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    11101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    11161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    11221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    11281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    11341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    11401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    11461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    11521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    11581 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    11641 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    11701 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    11761 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    11821 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcaga
    11881 attaaccctc actaaaggga
//
