LOCUS       sequence-366938-        8988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the sg83 helixCAM
            and constitutive expression of an identifying fluorescent protein 
            (iRFP670). Flanked with PiggyBac sites for easy line construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8988)
  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 8988)
  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_sg83-PDGFR_EF1a_iRFP670
FEATURES             Location/Qualifiers
     source          1..8988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..222
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    267..291
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     291..350
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             360..377
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             504..533
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             537..683
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(694..711)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    700..907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        984..1195
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1208..1476
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1510..2253
                     /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             2254..2307
                     /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             2308..3240
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             3241..3294
                     /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             3295..3894
                     /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(3295..3314)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3791..3811
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3927..4048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4018..4037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4811..4845
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5336..5352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5336..5352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5349..5371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5360..5376
                     /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(5384..5390,5391..5408,5409..5414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6497..7288,7289..7357))
                     /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     7120..7139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7358..7462)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7488..7943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7625..7646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7837..7856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8070..8092
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8084..8101
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8085..8101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   8231..8293
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        join(8742..8988,1..132)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    8750..8768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8786..8804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8822..8840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8858..8876
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8894..8912
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8930..8948
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8966..8984
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc
       61 agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg taccacttcc
      121 taccctcgta aaagtctaga gctagcgaat tcgaagccac caactggcag gtgcgtatgt
      181 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      241 atcgcctgga gaattggcta gcatcaacaa gtttgtacaa aaaagttggc atggagacag
      301 acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt gacggaagcc
      361 atcaccacca tcatcacggt gcatctcctg aagacaagaa cgaggaacta aaatccaaga
      421 ttgccgaatt gaaaaataag aatgaagcgt tagagcagaa gattgaacaa ctgaagcagg
      481 agattgctaa cttggagaaa gacgaacaaa aactcatctc agaagaggat ctgaatgctg
      541 tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt aaggtggtgg
      601 tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc atcctcatca
      661 tgctttggca gaagaagcca cgttaacggt acccctcgac tgtgccttct agttgccagc
      721 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
      781 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
      841 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
      901 ctggggatgc ggtgggctct tcgaatttaa atcggatcca gctgaatcta agtggatctg
      961 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     1021 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     1081 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     1141 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     1201 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     1261 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     1321 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     1381 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     1441 ctttgtttcg ttttctgttc tgcgccgtta cagatctaag ctgtgaccgg cgcctactct
     1501 agagccgcca tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc
     1561 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     1621 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     1681 gagatgctgg acaggcatca tacccactcc tgccccctgg aaggcgagtc atggcaagac
     1741 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
     1801 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     1861 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     1921 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     1981 agagagacac ctaccaccga ttctatgccc ccacttctga aacaagcaat tgagctgttc
     2041 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     2101 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
     2161 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     2221 cttgacgatt ttgaccttga catgctcccc ggggagggca gaggaagtct tctaacatgc
     2281 ggtgacgtgg aggagaatcc cggccctatg gcgcgtaagg tcgatctcac ctcctgcgat
     2341 cgcgagccga tccacatccc cggcagcatt cagccgtgcg gctgcctgct agcctgcgac
     2401 gcgcaggcgg tgcggatcac gcgcattacg gaaaatgccg gcgcgttctt tggacgcgaa
     2461 actccgcggg tcggtgagct actcgccgat tacttcggcg agaccgaagc ccatgcgctg
     2521 cgcaacgcac tggcgcagtc ctccgatcca aagcgaccgg cgctgatctt cggttggcgc
     2581 gacggcctga ccggccgcac cttcgacatc tcactgcatc gccatgacgg tacatcgatc
     2641 atcgagttcg agcctgcggc ggccgaacag gccgacaatc cgctgcggct gacgcggcag
     2701 atcatcgcgc gcaccaaaga actgaagtcg ctcgaagaga tggccgcacg ggtgccgcgc
     2761 tatctgcagg cgatgctcgg ctatcaccgc gtgatgttgt accgcttcgc ggacgacggc
     2821 tccgggatgg tgatcggcga ggcgaagcgc agcgacctcg agagctttct cggtcagcac
     2881 tttccggcgt cgctggtccc gcagcaggcg cggctactgt acttgaagaa cgcgatccgc
     2941 gtggtctcgg attcgcgcgg catcagcagc cggatcgtgc ccgagcacga cgcctccggc
     3001 gccgcgctcg atctgtcgtt cgcgcacctg cgcagcatct cgccctgcca tctcgaattt
     3061 ctgcggaaca tgggcgtcag cgcctcgatg tcgctgtcga tcatcattga cggcacgcta
     3121 tggggattga tcatctgtca tcattacgag ccgcgtgccg tgccgatggc gcagcgcgtc
     3181 gcggccgaaa tgttcgccga cttcttatcg ctgcacttca ccgccgccca ccaccaacgc
     3241 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc
     3301 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3361 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3421 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3481 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3541 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3601 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3661 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3721 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3781 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3841 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa
     3901 cctctggatt acaaaatttg tgggtaaact tgtttattgc agcttataat ggttacaaat
     3961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4021 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat
     4081 tagtctatca gaagctcatc tggtctccct tccgggggac aagacatccc tgtttaatat
     4141 ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc
     4201 agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc
     4261 cccggaattg actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc
     4321 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc
     4381 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg
     4441 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa
     4501 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat
     4561 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     4621 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     4681 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     4741 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     4801 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat
     4861 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg
     4921 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag
     4981 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg
     5041 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg
     5101 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc
     5161 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac
     5221 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag
     5281 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat
     5341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6781 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7081 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     7501 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7561 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7621 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7681 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7741 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7801 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7861 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7921 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     7981 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8041 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     8101 tgagcgcgcc tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc
     8161 aaatgtgttt tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca
     8221 gctagattaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgtgta
     8281 aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga
     8341 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
     8401 tatgtttatt tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa
     8461 acttttatga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
     8521 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
     8581 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
     8641 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
     8701 ggcctccacg gccactagtc tttcgtcttc aagaattcct cgagtttact ccctatcagt
     8761 gatagagaac gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta
     8821 ctccctatca gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt
     8881 atgaccagtt tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt
     8941 gatagagaac gtatatccag tttactccct atcagtgata gagaacgt
//
