LOCUS       sequence-366935-        8772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the sg88 helixCAM
            and constitutive expression of an identifying fluorescent protein 
            (eBFP2). Flanked with PiggyBac sites for easy line construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8772)
  AUTHORS   Chao G, Wannier TM, Gutierrez C, Borders NC, Appleton E, Chadha A, 
            Lebar T, Church GM
  TITLE     helixCAM: A platform for programmable cellular assembly in bacteria 
            and human cells.
  JOURNAL   Cell. 2022 Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012.
  PUBMED    36055250
REFERENCE   2  (bases 1 to 8772)
  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: "Cell. 2022 
            Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012."
COMMENT     Sequence Label: PB_TRE_sg88-PDGFR_EF1a_eBFP2
FEATURES             Location/Qualifiers
     source          1..8772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1519..1974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1656..1677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1868..1887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2101..2123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2115..2132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2116..2132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2262..2324
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2773..3151
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2781..2799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2817..2835
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2853..2871
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2889..2907
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2925..2943
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2961..2979
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2997..3015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3203..3241
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    3286..3310
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     3310..3369
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             3379..3396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3523..3552
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3556..3702
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(3713..3730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3719..3926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4003..4214
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4227..4495
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4529..5272
                     /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             5273..5326
                     /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(5327..5329,5330..5332,5333..6043)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(5372..5393)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6044..6097
                     /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             6098..6697
                     /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(6098..6117)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6594..6614
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6730..6851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6767..6786)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6821..6840
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7614..7648
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8139..8155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8139..8155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8152..8174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8163..8179
                     /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(8187..8193,8194..8211,8212..8217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8232..8253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8370..8387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8541..8772,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8621..8640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1561 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1621 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     1681 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1741 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1801 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     1861 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     1921 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     1981 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2041 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2101 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc ctcgttcatt cacgtttttg
     2161 aacccgtgga ggacgggcag actcgcggtg caaatgtgtt ttacagcgtg atggagcaga
     2221 tgaagatgct cgacacgctg cagaacacgc agctagatta accctagaaa gataatcata
     2281 ttgtgacgta cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt
     2341 atatcgaggt ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca
     2401 tactaataat aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa
     2461 aaactcaaaa tttcttctat aaagtaacaa aacttttatg agggacagcc cccccccaaa
     2521 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     2581 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     2641 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     2701 cctgcagaca cctgggggga tacggggaaa aggcctccac ggccactagt ctttcgtctt
     2761 caagaattcc tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc
     2821 tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag
     2881 gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag
     2941 agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc
     3001 tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga
     3061 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc
     3121 aactttccgt accacttcct accctcgtaa aggtctagag ctagcgaatt cgaagccacc
     3181 aactggcagg tgcgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc
     3241 tcgtttagtg aaccgtcaga tcgcctggag aattggctag catcaacaag tttgtacaaa
     3301 aaagttggca tggagacaga cacactcctg ctatgggtac tgctgctctg ggttccaggt
     3361 tccactggtg acggaagcca tcaccaccat catcacggtg caagtccgga ggataaacta
     3421 gccacgatta agaacaaact ggccgaaata aagaatgagc tagcgagaaa taagaaagaa
     3481 ctagcaaata tagaaagaga actagcaaag aacgagagag acgaacaaaa actcatctca
     3541 gaagaggatc tgaatgctgt gggccaggac acgcaggagg tcatcgtggt gccacactcc
     3601 ttgcccttta aggtggtggt gatctcagcc atcctggccc tggtggtgct caccatcatc
     3661 tcccttatca tcctcatcat gctttggcag aagaagccac gttaacggta cccctcgact
     3721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3901 gaagacaata gcaggcatgc tggggatgcg gtgggctctt cgaatttaaa tcggatccag
     3961 ctgaatctaa gtggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     4021 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     4081 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     4141 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     4201 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     4261 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     4321 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     4381 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     4441 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatctaagc
     4501 tgtgaccggc gcctactcta gagccgccat gtctagactg gacaagagca aagtcataaa
     4561 ctctgctctg gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     4621 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     4681 gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccactcct gccccctgga
     4741 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct
     4801 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     4861 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     4921 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     4981 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgaa
     5041 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     5101 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc
     5161 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
     5221 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg gggagggcag
     5281 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatgg tgagcaaggg
     5341 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5401 ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     5461 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     5521 gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     5581 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5641 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctagtga accgcatcga
     5701 gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5761 cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca tcaaggtgaa
     5821 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     5881 gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc tgagcaccca
     5941 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttccg
     6001 caccgccgcc gggatcactc tcggcatgga cgagctgtac aaggagggca gaggaagtct
     6061 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt
     6121 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     6181 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     6241 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     6301 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     6361 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     6421 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     6481 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     6541 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     6601 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     6661 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg attacaaaat
     6721 ttgtgggtaa acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6781 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6841 aatgtatctt atcatgtctg gaattgactc aaatgatgtc aattagtcta tcagaagctc
     6901 atctggtctc ccttccgggg gacaagacat ccctgtttaa tatttaaaca gcagtgttcc
     6961 caaactgggt tcttatatcc cttgctctgg tcaaccaggt tgcagggttt cctgtcctca
     7021 caggaacgaa gtccctaaag aaacagtggc agccaggttt agccccggaa ttgactggat
     7081 tcctttttta gggcccattg gtatggcttt ttccccgtat ccccccaggt gtctgcaggc
     7141 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     7201 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     7261 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     7321 ttgggggggg gctgtccctg atatctataa caagaaaata tatatataat aagttatcac
     7381 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     7441 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     7501 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     7561 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     7621 caattttacg cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg
     7681 gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga
     7741 agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg
     7801 ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc
     7861 catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg
     7921 gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac
     7981 cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg
     8041 atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc
     8101 ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     8161 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     8221 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     8281 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     8341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     8401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8761 tcggtgtagg tc
//
