LOCUS       sequence-320049-        8415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible overexpression of human SPI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  AUTHORS   Fazeli PK, Zhang Y, O'Keefe J, Pesaresi T, Lun M, Lawney B, 
            Steinhauser ML
  TITLE     Prolonged fasting drives a program of metabolic inflammation in 
            human adipose tissue.
  JOURNAL   Mol Metab. 2020 Sep 28;42:101082. doi: 10.1016/j.molmet.2020.101082.
  PUBMED    32992039
REFERENCE   2  (bases 1 to 8415)
  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: "Mol 
            Metab."; date: "2020-09-28"; volume: "42:101082. doi"; pages: " 
            10.1016/j.molmet.2020.101082"
COMMENT     Sequence Label: pCW-SPI1
FEATURES             Location/Qualifiers
     source          1..8415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..118
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        172..486
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    180..198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    251..269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    287..305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    323..341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    358..376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     459..483
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1317..1827
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1746..1764
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1837..2433
                     /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(1837..1856)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2333..2353
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2434..2487
                     /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             join(2488..3105,3106..3111,3112..3147,3148..3150,3151..3186,
                     3187..3189,3190..3225,3226..3234)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3274..3294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3692..3703)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3908..4141
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4213..4334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4250..4269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4304..4323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4374..4509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4509..4580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4581..4649,4650..5441)
                     /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(4799..4818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5612..6200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6354..6371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6488..6509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6524..6529,6530..6547,6548..6554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6562..6578
                     /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     6567..6589
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6586..6602
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6586..6602
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6621..6641
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6623..6641
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6669..6895
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6896..7076
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7123..7248
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7741..7974
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8159..8203
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
       61 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      121 ggtttattac agggacagca gagatccact ttggccgcga atcgatatgt cgagtttact
      181 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
      241 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      301 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      361 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      421 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      481 atcgcctgga gaattggcca ccatgttaca ggcgtgcaaa atggaagggt ttcccctcgt
      541 cccccctcca tcagaagacc tggtgcccta tgacacggat ctataccaac gccaaacgca
      601 cgagtattac ccctatctca gcagtgatgg ggagagccat agcgaccatt actgggactt
      661 ccacccccac cacgtgcaca gcgagttcga gagcttcgcc gagaacaact tcacggagct
      721 ccagagcgtg cagcccccgc agctgcagca gctctaccgc cacatggagc tggagcagat
      781 gcacgtcctc gataccccca tggtgccacc ccatcccagt cttggccacc aggtctccta
      841 cctgccccgg atgtgcctcc agtacccatc cctgtcccca gcccagccca gctcagatga
      901 ggaggagggc gagcggcaga gccccccact ggaggtgtct gacggcgagg cggatggcct
      961 ggagcccggg cctgggctcc tgcctgggga gacaggcagc aagaagaaga tccgcctgta
     1021 ccagttcctg ttggacctgc tccgcagcgg cgacatgaag gacagcatct ggtgggtgga
     1081 caaggacaag ggcaccttcc agttctcgtc caagcacaag gaggcgctgg cgcaccgctg
     1141 gggcatccag aagggcaacc gcaagaagat gacctaccag aagatggcgc gcgcgctgcg
     1201 caactacggc aagacgggcg aggtcaagaa ggtgaagaag aagctcacct accagttcag
     1261 cggcgaagtg ctgggccgcg ggggcctggc cgagcggcgc cacccgcccc actgacgggg
     1321 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
     1381 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
     1441 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
     1501 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
     1561 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
     1621 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
     1681 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
     1741 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
     1801 gaccgaatca ccgacctctc tccccagcaa ttcaccatga ccgagtacaa gcccacggtg
     1861 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     1921 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     1981 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     2041 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     2101 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     2161 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     2221 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     2281 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     2341 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     2401 cgcacctggt gcatgacccg caagcccggt gccgaaggta gaggttctct cctcacttgt
     2461 ggtgatgttg aagaaaaccc tggtccaatg tctagactgg acaagagcaa agtcataaac
     2521 ggagctctgg aattactcaa tggtgtcggt atcgaaggcc tgacgacaag gaaactcgct
     2581 caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg
     2641 ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa
     2701 ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cataccgctg tgctctcctc
     2761 tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa
     2821 accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg
     2881 tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat
     2941 caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga
     3001 caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa
     3061 ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc gaccgacgcc
     3121 cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat
     3181 atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt
     3241 aaggatcgat ccaagatatc gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3301 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3361 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3421 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3481 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3541 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3601 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3661 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3721 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3781 tcggatctcc ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg actagaggta
     3841 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     3901 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     3961 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4021 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4081 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4141 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     4201 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4321 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4561 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4621 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4681 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4741 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4801 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4861 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4921 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4981 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5041 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5101 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5161 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5221 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5281 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5341 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5401 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6361 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6481 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6541 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6601 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     6661 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     6721 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     6781 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     6841 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     6901 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6961 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7021 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     7081 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     7141 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     7201 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     7261 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     7321 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     7381 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     7441 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     7501 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     7561 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     7621 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     7681 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     7741 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     7801 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     7861 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     7921 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     7981 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     8041 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     8101 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     8161 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     8221 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     8281 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     8341 atcgtttcag acccacctcc caaccccgag gggacaattc tcgacctcga gacaaatggc
     8401 agtattcatc cacaa
//
