LOCUS       sequence-354927-       10374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10374)
  AUTHORS   Jung J, Chang Y, Jin G, Lian X, Bao X
  TITLE     Temporal Expression of Transcription Factor ID2 Improves Natural 
            Killer Cell Differentiation from Human Pluripotent Stem Cells.
  JOURNAL   ACS Synth Biol. 2022 May 24. doi: 10.1021/acssynbio.2c00017.
  PUBMED    35608547
REFERENCE   2  (bases 1 to 10374)
  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: "ACS Synth 
            Biol. 2022 May 24. doi: 10.1021/acssynbio.2c00017."
COMMENT     Sequence Label: AAVS1-PuroR Xlone eGFP
FEATURES             Location/Qualifiers
     source          1..10374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    77..913
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    987..1008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1023..1028,1029..1046,1047..1053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1061..1077
                     /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     1085..1101
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1444..1463
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1462..1517
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1563..1582
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1692..2168
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1950..1969
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2212..2958)
                     /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             complement(2959..2982)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2982..2991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3011..3222)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        3264..3631
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    3272..3290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3308..3326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3344..3362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3380..3398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3416..3434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3452..3470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3488..3506
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(3638..3640,3641..3643,3644..4354)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3638..3640,3641..3643,3644..4354)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3683..3704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3944..3963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4291..4312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4544..4678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4622..4641
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    4703..4927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4916..4933
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4965..5564)
                     /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(5048..5068)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5545..5564
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(5574..5627)
                     /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"
     misc_feature    5651..5676
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    5683..6486
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(6525..6543)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6564..6580)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6588..6604
                     /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(6612..6618,6619..6636,6637..6642))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6657..6678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6795..6812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6966..7554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7046..7065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7678..7746,7747..8538)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7896..7915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(8899..8918)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9513..9532
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(9935..10237)
                     /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     complement(10037..10056)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     join(10360..10374,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(10374,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg aatttagcgg
       61 ccgcgaattc gcccttagag cagagccagg aacccctgta gggaaggggc aggagagcca
      121 ggggcatgag atggtggacg aggaaggggg acagggaagc ctgagcgcct ctcctgggct
      181 tgccaaggac tcaaacccag aagcccagag cagggcctta gggaagcggg accctgctct
      241 gggcggagga atatgtccca gatagcactg gggactcttt aaggaaagaa ggatggagaa
      301 agagaaaggg agtagaggcg gccacgacct ggtgaacacc taggacgcac cattctcaca
      361 aagggagttt tccacacgga cacccccctc ctcaccacag ccctgccagg acggggctgg
      421 ctactggcct tatctcacag gtaaaactga cgcacggagg aacaatataa attggggact
      481 agaaaggtga agagccaaag ttagaactca ggaccaactt attctgattt tgtttttcca
      541 aactgcttct cctcttggga agtgtaagga agctgcagca ccaggatcag tgaaacgcac
      601 cagacagccg cgtcagagca gctcaggttc tgggagaggg tagcgcaggg tggccactga
      661 gaaccgggca ggtcacgcat cccccccttc cctcccaccc cctgccaagc tctccctccc
      721 aggatcctct ctggctccat cgtaagcaaa ccttagaggt tctggcaagg agagagatgg
      781 ctccaggaaa tgggggtgtg tcaccagata aggaatctgc ctaacaggag gtgggggtta
      841 gacccaatat caggagacta ggaaggagga ggcctaagga tggggctttt ctgtcaccaa
      901 tcctgtccct agtaaagctt agtactgtcg atccgctggc acgacaggtt tcccgactgg
      961 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1021 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1081 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     1141 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     1201 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     1261 tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg
     1321 gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat
     1381 atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     1441 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     1501 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     1561 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     1621 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattcacgcg cctggttacc
     1681 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
     1741 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
     1801 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
     1861 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
     1921 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
     1981 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
     2041 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
     2101 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
     2161 tgccacccgt agatctctcg agcagcgctc gatgatatcg aattcacgcg tttacccggg
     2221 gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc
     2281 gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt cggtcggccc
     2341 gccgctttcg cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag
     2401 gaaggcaggt tcggctccct gccggtcgaa cagctcaatt gcttgtttca gaagtggggg
     2461 catagaatcg gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc
     2521 caatacgcag cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag
     2581 ggagaagcct tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc
     2641 tgttgggcgg gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg
     2701 gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagga
     2761 gtgggtatga tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt
     2821 cttcacgtgc cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt
     2881 cgtcaggcct tcgataccga ctccattgag taattccaga gcagagttta tgactttgct
     2941 cttgtccagt ctagacatct tatcgtcatc gtctttgtaa tccatggtgg cggatcccgc
     3001 gtcacgacac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     3061 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     3121 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
     3181 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     3241 cactcgagtg gaattatcac ctcgagttta ctccctatca gtgatagaga acgtatgaag
     3301 agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga
     3361 gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct
     3421 atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc
     3481 agtttactcc ctatcagtga tagagaacgt ataagctttg cttatgtaaa ccagggcgcc
     3541 tataaaagag tgctgatttt ttgagtaaac ttcaattcca caacactttt gtcttatacc
     3601 aactttccgt accacttcct accctcgtaa aggtaccatg gtgagcaagg gcgaggagct
     3661 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3721 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3781 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3841 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3901 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3961 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4021 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4081 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     4141 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4201 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4261 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4321 cgggatcact ctcggcatgg acgagctgta caaggaattc taaactagta gaccacctcc
     4381 cctgcgagct aagctggaca gccaatgacg ggtaagagag tgacattttt cactaaccta
     4441 agacaggagg gccgtcagag ctactgccta atccaaagac gggtaaaagt gataaaaatg
     4501 tatcactcca acctaagaca ggcgcagctt ccgagggatt tgagatccag acatgataag
     4561 atacattgat gagtttggac aaaccaaaac tagaatgcag tgaaaaaaat gccttatttg
     4621 tgaaatttgt gatgctattg ccttatttgt aaccattata agctgcaata aacaagttct
     4681 agtcaataat caatgtcgag acccatagag cccaccgcat ccccagcatg cctgctattg
     4741 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     4801 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     4861 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     4921 ggcacagtcg aggctgatca gcgggtttaa acgggccctc tagatcaggc accgggcttg
     4981 cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag
     5041 ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc
     5101 ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc
     5161 tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc
     5221 ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc
     5281 atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag
     5341 accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc
     5401 gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg
     5461 tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg
     5521 tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatctcgag cctagggccg
     5581 ggattctcct ccacgtcacc gcatgttaga agacttcctc tgccctctcc gctgccagat
     5641 ctctcgaggc cctgtgggag gaagagaaga ggtcagaagc ttgccccact gtggggtgga
     5701 ggggacagat aaaagtaccc agaaccagag ccacattaac cggccctggg aatataaggt
     5761 ggtcccagct cggggacaca ggatccctgg aggcagcaaa catgctgtcc tgaagtggac
     5821 ataggggccc gggttggagg aagaagacta gctgagctct cggacccctg gaagatgcca
     5881 tgacaggggg ctggaagagc tagcacagac tagagaggta aggggggtag gggagctgcc
     5941 caaatgaaag gagtgagagg tgacccgaat ccacaggaga acggggtgtc caggcaaaga
     6001 aagcaagagg atggagaggt ggctaaagcc agggagacgg ggtactttgg ggttgtccag
     6061 aaaaacggtg atgatgcagg cctacaagaa ggggaggcgg gacgcaaggg agacatccgt
     6121 cggagaaggc catcctaaga aacgagagat ggcacaggcc ccagaaggag aaggaaaagg
     6181 gaacccagcg agtgaagacg gcatggggtt gggtgaggga ggagagatgc ccggagagga
     6241 cccagacacg gggaggatcc gctcagagga catcacgtgg tgcagcgccg agaaggaagt
     6301 gctccggaaa gagcatcctt gggcagcaac acagcagaga gcaaggggaa gagggagtgg
     6361 aggaagacgg aacctgaagg aggcggcagg gaaggatctg ggccagccgt agaggtgacc
     6421 caggccacaa gctgcagaca gaaagcggca caggcccagg ggagagaatg ctggtcagag
     6481 aaagcaaagg gcgaattcgt ttaaacctgc aggactagct agtcccttta gtgagggtta
     6541 attctgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6601 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6661 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6721 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6781 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6841 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6901 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6961 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7021 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7081 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7141 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7201 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7261 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7321 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7381 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7441 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7501 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7561 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7621 ttggtcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7981 gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgagc
     8641 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8701 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8761 catcaggaaa ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc
     8821 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     8881 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     8941 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     9001 cggcaagcag gcatcgccat gcatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     9061 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     9121 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     9181 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     9241 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     9301 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     9361 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     9421 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     9481 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     9541 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     9601 atcctgtctc ttgatcagag cttgatcccc tgcgccatca gatccttggc ggcaagaaag
     9661 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
     9721 ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc
     9781 aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     9841 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgaaa aggatctagg
     9901 tgaagatcct ttttgataat ctcatgcctg acatttatat tccccagaac atcaggttaa
     9961 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
    10021 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
    10081 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
    10141 tcaccatccg tcgccccggc gtgtcaataa tatcactctg tacatccaca aacagacgat
    10201 aacggctctc tcttttatag gtgtaaacct taaactgccg tacgtatagg ctgcgcaact
    10261 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
    10321 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
