LOCUS       sequence-258114-       11444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  generation of Tet-On RPE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11444)
  AUTHORS   Ardeljan D, Steranka JP, Liu C, Li Z, Taylor MS, Payer LM, Gorbounov
            M, Sarnecki JS, Deshpande V, Hruban RH, Boeke JD, Fenyo D, Wu PH, 
            Smogorzewska A, Holland AJ, Burns KH
  TITLE     Cell fitness screens reveal a conflict between LINE-1 
            retrotransposition and DNA replication.
  JOURNAL   Nat Struct Mol Biol. 2020 Feb;27(2):168-178. doi: 
            10.1038/s41594-020-0372-1. Epub 2020 Feb 10.
  PUBMED    32042151
REFERENCE   2  (bases 1 to 11444)
  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"; doi: 
            "10.1038/s41594-020-0372-1"; journalName: "Nat Struct Mol Biol"; 
            date: "2020-02"; volume: "27"; issue: "2"; pages: "168-178"
COMMENT     Sequence Label: pDA093
FEATURES             Location/Qualifiers
     source          1..11444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..368
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    64..82
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    100..118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    135..153
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    171..189
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    207..225
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    278..296
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     341..365
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    5378..5499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5389..5408)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5443..5462
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             6222..6923
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(6355..6373)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6623..6642
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6817..6840
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             6933..6989
                     /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(6990..7607,7608..7613,7614..7649,7650..7652,7653..7688,
                     7689..7691,7692..7727,7728..7733)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             7740..7796
                     /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             7791..8591
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7851..7870)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8461..8480
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8646..8663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8652..8876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9202..9219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9373..9961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9453..9472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10132..10923,10924..10992))
                     /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     10755..10774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10993..11097)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcgactcct ctgcagaatg cggcgatgtt tcggtaaggg gtccgctatc tagacgagtt
       61 tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
      121 gatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
      181 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      241 atccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
      301 atgtcgatta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc
      361 agatcgcctg gagccaattc caactctttt gccttatacc tactttccgt accacttcct
      421 accctcgaaa ggcctctgat gggcaagaag cagaaccgca agaccggcaa cagcaagacc
      481 cagagcgcca gccccccccc caaggagcgc agcagcagcc ccgccaccga gcagagctgg
      541 atggagaacg acttcgacga gctgcgcgag gagggcttcc gccgcagcaa ctacagcgag
      601 ctgcgcgagg acatccagac caagggcaag gaggtggaga acttcgagaa gaacctggag
      661 gagtgcatca cccgcatcac caacaccgag aagtgcctga aggagctgat ggagctgaag
      721 accaaggccc gcgagctgcg cgaggagtgc cgcagcctgc gcagccgctg cgaccaattg
      781 gaggagcgcg tgagcgccat ggaggacgag atgaacgaga tgaagcgcga gggcaagttc
      841 cgcgagaagc gcatcaagcg caacgagcag agcctgcagg agatctggga ctacgtgaag
      901 cgccccaacc tgcgcctgat cggcgtgccc gagagcgacg tggagaacgg caccaagctg
      961 gagaacaccc tgcaggacat catccaggag aacttcccca acctggcccg ccaggccaac
     1021 gtgcagatcc aggagatcca gcgcaccccc cagcgctaca gcagccgccg cgccaccccc
     1081 cgccacatca tcgtgcgctt caccaaggtg gagatgaagg agaagatgct gcgcgccgcc
     1141 cgcgagaagg gccgcgtgac cctgaagggc aagcccatcc gtctcaccgc cgacctgagc
     1201 gccgagaccc tgcaggcccg ccgcgagtgg ggccccatct tcaacatcct gaaggagaag
     1261 aacttccagc cccgcatcag ctaccccgcc aagctgagct tcatcagcga gggcgagatc
     1321 aagtacttca tcgacaagca gatgctgcgc gacttcgtga ccacccgccc cgccctgaag
     1381 gagctgctga aggaggccct gaacatggag cgcaacaacc gctaccagcc cctgcagaac
     1441 cacgccaaga tgtaaagacc atcgagacta ggaagaaact gcatcaacta atgagcaaaa
     1501 tcaggcgcgc ccatcataat gaccggcagc aacagccaca tcaccatcct gaccctgaac
     1561 atcaacggcc tgaacagcgc catcaagcgc caccgcctgg ccagctggat caagagccag
     1621 gaccccagcg tgtgctgcat ccaggagacc cacctgacct gccgcgacac ccaccgcctg
     1681 aagatcaagg gctggcgcaa gatctaccag gccaacggca agcagaagaa ggccggcgtg
     1741 gccatcctgg tgagcgacaa gaccgacttc aagcccacca agatcaagcg cgacaaggag
     1801 ggccactaca tcatggtgaa gggcagcatc cagcaggagg agctgaccat cctgaacatc
     1861 tacgccccca acaccggtgc cccccgcttc atcaagcagg tgctgagcga cctgcagcgc
     1921 gacctggaca gccacaccct gatcatgggc gacttcaaca cccccctgag caccctggac
     1981 cgcagcaccc gccagaaggt gaacaaggac acccaggagc tgaacagcgc cctgcaccag
     2041 gccgacctga tcgacatcta ccgcaccctg caccccaaga gcaccgagta caccttcttc
     2101 agcgcccccc accacaccta cagcaagatc gaccacatcg tgggcagcaa ggccctgctg
     2161 agcaagtgca agcgcaccga gatcatcacc aactacctga gcgaccacag cgccatcaag
     2221 ctggagctgc gcatcaagaa cctgacccag agccgcagca ccacctggaa gctgaacaac
     2281 ctgctgctga acgactactg ggtgcacaac gagatgaagg ccgagatcaa gatgttcttc
     2341 gaaaccaacg agaacaagga caccacctac cagaacctgt gggacgcctt caaggccgtg
     2401 tgccgcggca agttcatcgc cctgaacgcc tacaagcgca agcaggagcg cagcaagatc
     2461 gacaccctga ccagccagct gaaggagctg gagaagcagg agcagaccca cagcaaggcc
     2521 agccgccgcc aggagatcac caagatccgc gccgagctga aggagatcga gacccagaag
     2581 accctgcaga agatcaacga gtcgcgaagc tggttcttcg agcgcatcaa caagatcgac
     2641 cgccccctgg cccgcctgat caagaagaag cgcgagaaga accagatcga caccatcaag
     2701 aacgacaagg gcgacatcac caccgacccc accgagatcc agaccaccat ccgcgagtac
     2761 tacaagcacc tgtacgccaa caagctggag aacctggagg agatggacac cttcctggac
     2821 acctacaccc tgccccgcct gaaccaggag gaggtggaga gcctgaaccg ccccatcacc
     2881 ggcagcgaga tcgtggccat catcaacagc ctgcccacca agaagagccc cggccccgac
     2941 ggcttcaccg ccgagttcta ccagcgctac aaggaggagc tggtgccctt cctgctgaag
     3001 ctgttccaaa gcatcgagaa ggagggcatc ctgcccaaca gcttctacga ggccagcatc
     3061 atcctgatcc ccaagcccgg ccgcgacacc accaagaagg agaacttccg ccccatcagc
     3121 ctgatgaaca tcgacgccaa gatcctgaac aagatcctgg ccaaccgcat ccagcagcac
     3181 atcaagaagc tgatccacca cgaccaggtg ggcttcatcc ccgggatgca gggctggttc
     3241 aacatccgca agagcatcaa cgtgatccag cacatcaacc gcgccaagga caagaaccac
     3301 atgatcatca gcatcgacgc cgagaaggcc ttcgacaaga tccagcagcc cttcatgctg
     3361 aagaccctga acaagctggg catcgacggc acctacttca agatcatccg cgccatctac
     3421 gacaagccca ccgccaacat catcctgaac ggccagaagc tggaggcctt ccccctgaag
     3481 accggcacgc gtcagggctg ccccctgagc cccctgctgt tcaacatcgt gctggaggtg
     3541 ctggcccgcg ccatccgcca ggagaaggag atcaagggca tccagctggg caaggaggag
     3601 gtgaagctga gcctgttcgc cgacgacatg atcgtgtacc tggagaaccc catcgtgagc
     3661 gcccagaacc tgctgaagct gatcagcaac ttcagcaagg tgagcggcta caagatcaac
     3721 gtgcagaaga gccaggcctt cctgtacacc aacaaccgcc agaccgagag ccagatcatg
     3781 ggcgagctgc ccttcaccat cgctagcaag cgcatcaagt acctgggcat ccagctgacc
     3841 cgcgacgtga aggacctgtt caaggagaac tacaagcccc tgctgaagga gatcaaggag
     3901 gagaccaaca agtggaagaa catcccctgc agctgggtgg gccgcatcaa catcgtgaag
     3961 atggccatcc tgcccaaggt gatctaccgc ttcaacgcca tccccatcaa gctgcccatg
     4021 accttcttca ccgagctgga gaagaccacc ctgaagttca tctggaacca gaagcgcgcc
     4081 cgcatcgcca agagcatcct gagccagaag aacaaggccg gcggcatcac cctgcccgac
     4141 ttcaagctgt actacaaggc caccgtgacc aagaccgcct ggtactggta ccagaaccgc
     4201 gatatcgacc agtggaaccg caccgagccc agcgagatca tgccccacat ctacaactac
     4261 ctgatcttcg acaagcccga gaagaacaag cagtggggca aggacagcct gttcaacaag
     4321 tggtgctggg agaactggct ggccatctgc cgcaagctga agctggaccc cttcctgacc
     4381 ccctacacca agatcaacag ccgctggatc aaggacctga acgtgaagcc caagaccatc
     4441 aagaccctgg aggagaacct gggcatcacc atccaggaca tcggcgtggg caaggacttc
     4501 atgagcaaga cccccaaggc catggccacc aaggacaaga tcgacaagtg ggacctgatc
     4561 aagctgaaga gcttctgcac cgccaaggag accaccatcc gcgtgaaccg ccagcccacc
     4621 acctgggaga agatcttcgc cacctacagc agcgacaagg gcctgatcag ccgcatctac
     4681 aacgagctga agcagatcta caagaagaag accaacaacc ccatcaagaa gtgggccaag
     4741 gacatgaacc gccacttcag caaggaggac atctacgccg ccaagaagca tatgaagaag
     4801 tgcagcagca gcctggccat ccgcgagatg cagatcaaga ccaccatgcg ctaccacctg
     4861 acccccgtgc gcatggccat catcaagaag agcggcaaca accgctgctg gcgcggctgc
     4921 ggcgagatcg gcaccctgct gcactgctgg tgggactgca agctggtgca gcccctgtgg
     4981 aagagcgtgt ggcgcttcct gcgcgacctg gagctggaga tccccttcga ccccgccatc
     5041 cccctgctgg gcatctaccc caacgagtac aagagctgct gctacaagga cacctgcacc
     5101 cgcatgttca tcgccgccct gttcaccatc gccaagacct ggaaccagcc caagtgcccc
     5161 accatgatcg attggatcaa gaagatgtgg cacatctaca ccatggagta ctacgccgcc
     5221 atcaagaacg acgagttcat cagcttcgtg ggcacctgga tgaagctgga gaccatcatc
     5281 ctgagcaagc tgagccagga gcagaagacc aagcaccgca tcttcagcct gatcggcggc
     5341 aactgatgtc aggccaagct tccatcgata gacatgataa gatacattga tgagtttgga
     5401 caaaccacaa caagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5461 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5521 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     5581 aaatgtggta cctaggcgag accctgtctc acaaaataaa gtaagcccgg actgagtgcg
     5641 gaaaggcggg cctggcgggt ctggtctccc catgcgggcc accagaggcc ctgcagcctt
     5701 cagtcgcttg aaggggtaat ggcgcttcca ctcacaaaca tggcggacag agcgtgtgaa
     5761 cgagatgaac agcccctcaa aaatatggcc gccgaggctg gacggccgtg ccccagcagc
     5821 accgcctccg cgccccacgt gatctctcgc cgggcacagc gctgaccgcg gaggtccaac
     5881 cggaagaatg tccggattgg acattcggaa gagggcccgc cttccctggg gaatctctgc
     5941 gcacgcgcag aacgcttcga ccaatgaaaa cacaggaagc cgtccgcgca accgcgttgc
     6001 gtcacttctg ccgcccctgt ttcaaggtat atagccgtag acggaacttc gcctttctct
     6061 cggccttagc gccatttttt tgggtgagtg ttttttggtt cctgcgttgg gattccgtgt
     6121 acaatccata gacatctgac ctcggcactt agcatcatca cagcaaacta actgtagcct
     6181 ttctctcttt ccctgtagaa acctctgcac ctgaggccac catgttgagc aagggcgagg
     6241 aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccatg
     6301 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     6361 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     6421 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     6481 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     6541 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     6601 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gacaatgggc tgggaggcct
     6661 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     6721 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     6781 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     6841 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt
     6901 acggcatgga cgagctgtac aagggcagtg gagctactaa cttcagcctg ctgaagcagg
     6961 ctggtgacgt cgaggagaat cctggcccca tgtctagact ggacaagagc aaagtcataa
     7021 acggcgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     7081 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     7141 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     7201 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc
     7261 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     7321 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     7381 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     7441 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     7501 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     7561 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccggccgacg
     7621 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     7681 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtccggcg
     7741 ctactaactt cagcctgctg aagcaggctg gtgacgtcga ggagaatcct ggtcccatga
     7801 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     7861 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     7921 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     7981 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     8041 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     8101 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     8161 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     8221 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     8281 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     8341 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     8401 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     8461 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     8521 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     8581 agttcttctg attcgaaggc ctgtcgtgaa gcttggggat caattctcta gagctcgctg
     8641 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     8701 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     8761 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     8821 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     8881 tgaggcggaa agaaccagct gggcctacta gctactcggg accccttacc gaaacatcgc
     8941 cgcattctgc agaggagtcg agtgtatgta aacttctgac ccactgggaa tgtgatgaaa
     9001 gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc acattcttaa
     9061 aataaagtgg tgatcctaac tgacctaaga cagggaattt ttactaggat taaatgtcag
     9121 gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac ttccgacttc
     9181 aactgtatag ggatccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     9241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     9301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     9361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     9421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     9481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     9541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     9601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     9661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     9721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     9781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     9841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     9901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     9961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    10021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    10081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    10141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    10201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    10261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    10321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    10381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    10441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    10501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    10561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    10621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    10681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    10801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    10861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    10921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    10981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    11041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    11101 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
    11161 aataccgcat caggaaattg taagcgttaa tattttgtta aaattggatc cctatacagt
    11221 tgaagtcgga agtttacata cacttaagtt ggagtcatta aaactcgttt ttcaactact
    11281 ccacaaattt cttgttaaca aacaatagtt ttggcaagtc agttaggaca tctactttgt
    11341 gcatgacaca agtcattttt ccaacaattg tttacagaca gattatttca cttataattc
    11401 actgtatcac aattccagtg ggtcagaagt ttacatacac taag
//
