LOCUS       sequence-354943-        9146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tunable and temporal expression control of CCND2, luciferase and 
            nuclear eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9146)
  AUTHORS   Joshi J, Xu B, Rubart M, Chang Y, Bao X, Chaliki HP, Scott LR, Zhu W
  TITLE     Optogenetic Control of Engrafted Human Induced Pluripotent Stem 
            Cell-Derived Cardiomyocytes in Live Mice: A Proof-of-Concept Study.
  JOURNAL   Cells. 2022 Mar 10;11(6). pii: cells11060951. doi: 
            10.3390/cells11060951.
  PUBMED    35326403
REFERENCE   2  (bases 1 to 9146)
  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: "Cells."; 
            date: "2022-03-10"; volume: "11(6). pii: cells11060951. doi"; pages:
            " 10.3390/cells11060951"
COMMENT     Sequence Label: XLone-Puro CCND2-T2A-luciferase-P2A-eGFP
FEATURES             Location/Qualifiers
     source          1..9146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1017..1079
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1255..1336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1357..1956)
                     /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(1440..1460)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1937..1956
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(1966..2019)
                     /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             complement(2029..2772)
                     /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(2773..2796)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2796..2805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2825..3036)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        3078..3445
                     /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    3086..3104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3122..3140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3158..3176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3194..3212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3230..3248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3266..3284
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3302..3320
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      3452..3461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4334..4387
                     /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             4388..6037
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             6053..6109
                     /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             join(6110..6112,6113..6115,6116..6826)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6110..6112,6113..6115,6116..6826)
                     /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(6155..6176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6416..6435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6763..6784
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6827..6847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7066..7200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   7476..7510
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7521..7538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7521..7537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7530..7552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7746..7765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7865..7887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7925..7943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8011..8115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8116..8184,8185..8976)
                     /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(8334..8353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaaggtcg acttaaccct
     1021 agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat tgacgcatgt
     1081 gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt aagttttatt
     1141 atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg tttatttatt
     1201 tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttagcagtg
     1261 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1321 ctgcaataaa caagttaact aaacgggccc tctagatcag gcaccgggct tgcgggtcat
     1381 gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg
     1441 ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg
     1501 ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc
     1561 gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag
     1621 gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg
     1681 gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac
     1741 cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa
     1801 gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt
     1861 ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg
     1921 ggtggcgagg cgcaccgtgg gcttgtactc ggtcatctcg agcctagggc cgggattctc
     1981 ctccacgtca ccgcatgtta gaagacttcc tctgccctct ccgctgcccc cggggagcat
     2041 gtcaaggtca aaatcgtcaa gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag
     2101 ggcatcggct gggagcatgt ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct
     2161 ttcgcacttt agctgtttct ccaggccaca tatgattagt tccaggccga aaaggaaggc
     2221 aggttcggct ccctgccggt cgaacagctc aattgcttgt ttcagaagtg ggggcataga
     2281 atcggtggta ggtgtctctc tttcctcttt tgctacttga tgctcctgtt cctccaatac
     2341 gcagcccagt gtaaagtggc ccacggcgga cagagcgtac agtgcgttct ccagggagaa
     2401 gccttgctga cacaggaacg cgagctgatt ttccagggtt tcgtactgtt tctctgttgg
     2461 gcgggtgccg agatgcactt tagccccgtc gcgatgtgag aggagagcac agcggtatga
     2521 cttggcgttg ttccgcagaa agtcttgcca tgactcgcct tccagggggc aggagtgggt
     2581 atgatgcctg tccagcatct cgattggcag ggcatcgagc agggcccgct tgttcttcac
     2641 gtgccagtac agggtaggct gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag
     2701 gccttcgata ccgactccat tgagtaattc cagagcagag tttatgactt tgctcttgtc
     2761 cagtctagac atcttatcgt catcgtcttt gtaatccatg gtggcggatc ccgcgtcacg
     2821 acacctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
     2881 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
     2941 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
     3001 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagccactcg
     3061 agtggaatta tcacctcgag tttactccct atcagtgata gagaacgtat gaagagttta
     3121 ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt
     3181 ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt
     3241 gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat atccagttta
     3301 ctccctatca gtgatagaga acgtataagc tttgcttatg taaaccaggg cgcctataaa
     3361 agagtgctga ttttttgagt aaacttcaat tccacaacac ttttgtctta taccaacttt
     3421 ccgtaccact tcctaccctc gtaaaggtac cgccaccatg gagctgctgt gccacgaggt
     3481 ggacccggtc cgcagggccg tgcgggaccg caacctgctc cgagacgacc gcgtcctgca
     3541 gaacctgctc accatcgagg agcgctacct tccgcagtgc tcctacttca agtgcgtgca
     3601 gaaggacatc caaccctaca tgcgcagaat ggtggccacc tggatgctgg aggtctgtga
     3661 ggaacagaag tgcgaagaag aggtcttccc tctggccatg aattacctgg accgtttctt
     3721 ggctggggtc ccgactccga agtcccatct gcaactcctg ggtgctgtct gcatgttcct
     3781 ggcctccaaa ctcaaagaga ccagcccgct gaccgcggag aagctgtgca tttacaccga
     3841 caactccatc aagcctcagg agctgctgga gtgggaactg gtggtgctgg ggaagttgaa
     3901 gtggaacctg gcagctgtca ctcctcatga cttcattgag cacatcttgc gcaagctgcc
     3961 ccagcagcgg gagaagctgt ctctgatccg caagcatgct cagaccttca ttgctctgtg
     4021 tgccacggac tttaagtttg ccatgtaccc accgtcgatg atcgcaactg gaagtgtggg
     4081 agcagccatc tgtgggctcc agcaggatga ggaagtgagc tcgctcactt gtgatgccct
     4141 gactgagctg ctggctaaga tcaccaacac agacgtggat tgtctcaaag cttgccagga
     4201 gcagattgag gcggtgctcc tcaatagcct gcagcagtac cgtcaggacc aacgtgacgg
     4261 atccaagtcg gaggatgaac tggaccaagc cagcacccct acagacgtgc gggatatcga
     4321 cctgggcagc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     4381 cggccctatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
     4441 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg
     4501 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
     4561 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
     4621 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
     4681 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
     4741 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
     4801 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     4861 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     4921 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     4981 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     5041 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     5101 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     5161 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     5221 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     5281 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     5341 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     5401 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     5461 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     5521 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     5581 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     5641 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     5701 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     5761 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     5821 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat
     5881 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     5941 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     6001 tctcattaag gccaagaagg gcggcaagat cgccgtggct agcggcagcg gcgccactaa
     6061 cttctccctg ttgaaacaag caggggatgt cgaagagaat cccgggccaa tggtgagcaa
     6121 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     6181 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     6241 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     6301 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     6361 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     6421 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     6481 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     6541 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     6601 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     6661 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     6721 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     6781 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagccaa agaagaagcg
     6841 taaggtataa gtacaagtaa gaattccgat catattcaat aacccactag tagaccacct
     6901 cccctgcgag ctaagctgga cagccaatga cgggtaagag agtgacattt ttcactaacc
     6961 taagacagga gggccgtcag agctactgcc taatccaaag acgggtaaaa gtgataaaaa
     7021 tgtatcactc caacctaaga caggcgcagc ttccgaggga tttgagatcc agacatgata
     7081 agatacattg atgagtttgg acaaaccaaa actagaatgc agtgaaaaaa atgccttatt
     7141 tgtgaaattt gtgatgctat tgccttattt gtaaccatta taagctgcaa taaacaagtt
     7201 tgatatctat aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat
     7261 aacaatataa ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc
     7321 attttgactc acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca
     7381 cgcctcacgg gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc
     7441 gctatttaga aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat
     7501 ctttctaggg ttaagaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     7561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     7621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     7681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     7741 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7801 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7861 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     7921 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     7981 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     8041 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     8101 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     8161 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     8221 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     8281 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     8341 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     8401 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     8461 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     8521 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     8581 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     8641 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     8701 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     8761 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     8821 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8881 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8941 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     9001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     9061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     9121 cccgtagaaa agatcaaagg atcttc
//
