LOCUS       Exported                8151 bp ds-DNA     circular SYN 26-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8151)
  AUTHORS   R van Ineveld
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 26, 2021 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8151
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    16..279
                     /label=Tol2
     misc_feature    280..285
                     /label=Bgl2_misc_feature
                     /note="note=""Bgl2"" - 

                     280..285"
     misc_feature    286..1448
                     /label=hEF1a promoter
     misc_feature    286..304
                     /label=N-terminus_misc_feature [*]
                     /note="note=""N-terminus"" - 1698..1721"
     misc_feature    298..304
                     /label=Kozak_misc_feature
                     /note="note=""Kozak"" - 

                     1715..1721"
     primer_bind     complement(646..661)
                     /label=BP406 Turq2 van F7 into miniTol-Y-ER-C_Rv
     promoter        1254..1448
                     /label=mouse phosphoglycerate kinase 1 promoter_PGK pr...
                     /note="note=""PGK promoter"" - note=""mouse 
                     phosphoglycerate kinase 1 promoter"" - 

                     2671..3170"
     primer_bind     1357..1376
                     /label=BP308 seq vanaf EF1a
     primer_bind     complement(1431..1498)
                     /label=BP489 EF1a ipv CMV  Rv
     primer_bind     complement(1457..1522)
                     /label=BP245 prepare pLV for project T Rev
     primer_bind     1500..1530
                     /label=BP326 Primer YPet into F7 Fw
     primer_bind     1505..1573
                     /label=BP306 mVenus into F7 Fw
     primer_bind     complement(1517..1533)
                     /label=BP094 Fusion PCR PGK-Puro 02 Rev
     primer_bind     1519..1573
                     /label=BP 219 EKAREVnls to pLV Fw
     primer_bind     1519..1558
                     /label=BP 225 EKAREVnls to pLV Fw
     primer_bind     1524..1571
                     /label=BP197 Flag-mNeon into F2 Fw
     primer_bind     1535..1558
                     /label=BP 226 EKAREVnls to E4 (or EF1a) Fw
     primer_bind     1544..1563
                     /label=BP335 Seq multi-acceptor Fw
     primer_bind     complement(1544..1563)
                     /label=BP336 Seq multi-acceptor Rev
     primer_bind     1554..1573
                     /label=BP460 Scarlet-I into K19 Fw
     primer_bind     1555..1576
                     /label=BP 050 Lgr5BAC_HA1_GFP_fw
     primer_bind     1555..1574
                     /label=BP317 Make Q2 PCR2 Fw
     primer_bind     1555..1573
                     /label=BP081 GFP in pCS2 Fw
     primer_bind     1555..1573
                     /label=BP323 mVenus into F11 Fw
     primer_bind     1555..1573
                     /label=BP 221 EKAREVnls to E4 (or EF1a) Fw
     primer_bind     1555..1573
                     /label=BP464 Scarlet-I into K9 PCR2 Fw
     primer_bind     1555..1572
                     /label=BP157 mTurq2-NES into C2 Fw
     primer_bind     1555..1571
                     /label=BP195 mNeon-CAAX into K5 Fw
     primer_bind     1555..1571
                     /label=BP185 mNeon into K1 Fw
     primer_bind     1555..1571
                     /label=BP458 Scarlet-I into K4 Fw
     primer_bind     1555..1571
                     /label=BP193 mNeon into CFP-CX Fw
     primer_bind     1555..1570
                     /label=BP468 Make TV22 PCR1 Fw
                     /note="(67 bp)"
     misc_feature    1556..2266
                     /label=YPet
     primer_bind     1556..1573
                     /label=BP201 GFP-CENP-B into K10 Fw
     primer_bind     1556..1573
                     /label=BP393 mCh-IRES to T24 Fw
     primer_bind     1556..1573
                     /label=BP401 Fragm2_Fw
     primer_bind     1556..1572
                     /label=BP159 mCherry-CAAX to C8-Y+C  Fw
     primer_bind     1623..1642
                     /label=BP 010 seq GFP Rev70
     primer_bind     complement(1630..1648)
                     /label=BP 053 GFP_HA1_control_rev
     primer_bind     complement(1677..1693)
                     /label=BP 009 seq GFP Rev120
     primer_bind     complement(1683..1706)
                     /label=BP360 Make Q8c Rev
     primer_bind     2095..2114
                     /label=BP 007 seq GFP Fw540
     primer_bind     complement(2261..2280)
                     /label=BP327 Primer YPet into F7 Rev
     misc_feature    2273..2437
                     /label=WW domain
     primer_bind     complement(2391..2408)
                     /label=BP334 Seq EKAREV from WW
     primer_bind     complement(2448..2467)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2448..2467)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2450..2665
                     /label=EV linker
     misc_feature    2450..2461
                     /label=Feature 23
     primer_bind     complement(2460..2485)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2460..2485)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2462..2473
                     /label=Feature 24
     misc_feature    2474..2479
                     /label=Feature 25
     misc_feature    2480..2491
                     /label=Feature 43
     primer_bind     complement(2490..2509)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2490..2509)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2492..2503
                     /label=Feature 32
     primer_bind     complement(2502..2527)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2502..2527)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2504..2521
                     /label=Feature 46
     primer_bind     complement(2520..2539)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2520..2539)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2522..2533
                     /label=Feature 33
     primer_bind     complement(2532..2551)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2532..2551)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2534..2545
                     /label=Feature 34
     primer_bind     complement(2544..2563)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2544..2563)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2546..2557
                     /label=Feature 35
     primer_bind     complement(2556..2581)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2556..2581)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2558..2569
                     /label=Feature 36
     misc_feature    2570..2587
                     /label=Feature 44
     primer_bind     complement(2586..2605)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2586..2605)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2588..2599
                     /label=Feature 37
     primer_bind     complement(2598..2623)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2598..2623)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2600..2617
                     /label=Feature 47
     primer_bind     complement(2616..2635)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2616..2635)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     misc_feature    2618..2629
                     /label=Feature 38
     misc_feature    2630..2641
                     /label=Feature 39
     misc_feature    2642..2653
                     /label=Feature 40
     primer_bind     complement(2652..2701)
                     /label=BP373 Mut F7 G414S Rev????????
     primer_bind     complement(2652..2701)
                     /label=BP375 Mut F7 S413P+G414S Rev????????
     primer_bind     2652..2701
                     /label=BP372 Mut F7 G414S Fw?????????
     primer_bind     2652..2701
                     /label=BP374 Mut F7 S413P+G414S Fw?????????
     misc_feature    2654..2665
                     /label=Feature 41
     primer_bind     complement(2670..2731)
                     /label=BP414 Mut F7 D423K, R419S Rv
     primer_bind     2670..2731
                     /label=BP413 Mut F7 D423K,R419S Fw
     primer_bind     complement(2673..2712)
                     /label=BP392 Mut F7-PW TA Rv
     primer_bind     2673..2712
                     /label=BP391 Mut F7-PW TA Fw
     primer_bind     complement(2673..2704)
                     /label=BP357 F7-P-mimetic E Rev
     primer_bind     complement(2673..2704)
                     /label=BP355 F7-P-mimetic D Rev
     primer_bind     complement(2675..2731)
                     /label=BP377 Mut F7 V422T Rv
     primer_bind     2675..2731
                     /label=BP376 Mut F7 V422T Fw
     misc_feature    2678..2731
                     /label=ERK Substrate
     primer_bind     2678..2731
                     /label=BP411 Mut F7 D423K Fw
     primer_bind     complement(2678..2704)
                     /label=BP224 Mutagenize (T-A) ERK sensor Rev
     primer_bind     2679..2730
                     /label=BP407 Mut F7 PW-to-W Fw
     primer_bind     2681..2735
                     /label=BP385 Mut F7 KK424,426PE Fw
     primer_bind     2681..2735
                     /label=BP383 Mut F7 K426E Fw
     primer_bind     2681..2735
                     /label=BP381 Mut F7 K426W Fw
     primer_bind     2681..2735
                     /label=BP379 Mut F7 KK424,426PW Fw
     primer_bind     2681..2735
                     /label=BP370 Mut F7 KK424,426PP Fw
     primer_bind     2681..2730
                     /label=BP366 Mut F7 K424P Fw?????? ???????????????
     primer_bind     2684..2735
                     /label=BP368 Mut F7 K426P Fw?????????
     primer_bind     2687..2735
                     /label=BP409 Mut F7 PW-to-P Fw
     misc_feature    2693..2695
                     /label=T-A
     primer_bind     2694..2740
                     /label=BP435 Mut F7 Dock F429D Fw
     primer_bind     2696..2746
                     /label=BP415 Mut F7 No Dock Fw
     CDS             2705..2707
                     /label=P
     CDS             2711..2713
                     /label=W
     primer_bind     2735..2754
                     /label=BP332 Primer Turq2 into F20 Fw
     primer_bind     2735..2754
                     /label=BP364 Make F22 Fw
     primer_bind     2740..2764
                     /label=BP421 Turq2 to YERC Frgm2 Fw
     CDS             2741..3451
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)Mutation K206A = monomeric to sticky
                     Mutation V224L = enhances dimerization sysynergistically 
                     with S208F of YPet (Vinkenborg. J.L. et al., 2007)
                     "
                     /label=Turquoise2 FRET-optimized
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFLTAAGITLGMDEL"
     primer_bind     2771..2795
                     /label=BP405 Turq2 van F7 into miniTol-Y-ER-C_Fw
     variation       2936..2938
                     /label=T65S , mTurq
     variation       2939..2941
                     /label=Y66W
     variation       2957..2959
                     /label=S72A
     variation       3128..3130
                     /label=G129D
     misc_feature    3179..3181
                     /label=I146F , mTurq2
     variation       3200..3202
                     /label=M153T
     variation       3230..3232
                     /label=V163A
     primer_bind     3257..3274
                     /label=BP345 Seq Fw vanaf mutated Turq
     primer_bind     complement(3257..3274)
                     /label=Primer 2
     variation       3266..3268
                     /label=S175G
     misc_feature    3359..3361
                     /label=K206A monomeric->sticky
     variation       3413..3415
                     /label=V224L enhances dimerization
     primer_bind     complement(3434..3452)
                     /label=BP422 Turq2 to YERC Frgm2 Rev
     misc_feature    3452..3456
                     /label=eCFP
     CDS             3458..3478
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3469..3501)
                     /label=BP 220 EKAREVnls to pLV Rev
     primer_bind     complement(3469..3491)
                     /label=BP 222 EKAREVnls to E4 (or EF1a) Rev
     primer_bind     complement(3509..3524)
                     /label=BP454 H2B-Neon to F80 Rv
     primer_bind     3534..3560
                     /label=BP295 make S1, polyA into E4, Fw
     promoter        complement(3534..3551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3545..3576
                     /label=BP499 polyA into TV22 Fw
     polyA_signal    3556..3690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3572..3598)
                     /label=BP456 Insulator into T30 Fw
     primer_bind     complement(3572..3598)
                     /label=BP466 pUNKI-insulator into K4 Fw
     primer_bind     3633..3656
                     /label=BP190 Seq Rev into TagGFP of Casper3
     primer_bind     complement(3725..3750)
                     /label=BP 043 PGK-Puro-pA Rv
     primer_bind     complement(3725..3750)
                     /label=BP077 PGK-Puro-pA Rv
     primer_bind     complement(3725..3750)
                     /label=BP500 polyA into TV22 Rv
     primer_bind     complement(3726..3751)
                     /label=BP475 Make TV26 PCR2 Rev
     primer_bind     complement(3727..3750)
                     /label=BP296 make S1, polyA into E4, Rev
     primer_bind     3727..3746
                     /label=BP067 Seq primer polyA Fw
     primer_bind     complement(3727..3746)
                     /label=BP066 Seq primer polyA Rev
     primer_bind     complement(3730..3755)
                     /label=bp341 TagRFP into Q3 Rev
     primer_bind     complement(3735..3755)
                     /label=BP439 Make F7-NES Rev
     primer_bind     3753..3768
                     /label=BP229 PGK-PuroDTK for T2 Fw
     primer_bind     3753..3768
                     /label=BP249 PGK-PruDTK for T2 Fw
     primer_bind     3753..3766
                     /label=BP 042 PGK-Puro-pA Fw
     primer_bind     3753..3766
                     /label=BP169 PGK:Puro w/o polyAis Fw
     primer_bind     3753..3766
                     /label=BP167 PGK:Puro w/o polyAis Fw
     promoter        3754..4253
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3793..3809)
                     /label=BP406 Turq2 van F7 into miniTol-Y-ER-C_Rv
     primer_bind     3906..3924
                     /label=Primer 3
     primer_bind     complement(4155..4165)
                     /label=BP435 Mut F7 Dock F429D Fw
     primer_bind     complement(4186..4205)
                     /label=BP120 PGK Seq Rev 430 (2)
     primer_bind     complement(4186..4205)
                     /label=BP120 PGK Seq Rev 430
     primer_bind     complement(4254..4273)
                     /label=BP484 CreER into TV26-EcoRI Rv
     primer_bind     complement(4254..4273)
                     /label=BP482 CreER into TV26 Rv
     promoter        4265..4312
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(4295..4322)
                     /label=BP240 prepare pLV for project T Rev
     primer_bind     complement(4295..4322)
                     /label=BP242 prepare pLV for project T Rev
     CDS             4331..4729
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     4519..4533
                     /label=BP334 Seq EKAREV from WW
     polyA_signal    4783..5007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    5029..5240
                     /label=Tol2
     primer_bind     5250..5267
                     /label=BP449 adapt miniTol for INS Rv
     promoter        complement(5276..5294)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5312..5328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5336..5352
                     /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(5360..5390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5387..5406)
                     /label=BP258 Seq pr in miniTol vector (T2) Rev
     protein_bind    5405..5426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5714..6302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6473..7333)
                     /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(6488..6507)
                     /label=BP 048 Amp_HA2_control_fw
     promoter        complement(7334..7438)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7516..7971)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8112..8128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(8135..8151,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(8147..8151,1..15)
                     /label=BP213 Mash3x into E7-gcamp Fw
ORIGIN
        1 gggcgaattg ggcccagagg tgtaaagtac ttgagtaatt ttacttgatt actgtactta
       61 agtattattt ttggggattt ttactttact tgagtacaat taaaaatcaa tacttttact
      121 tttacttaat tacatttttt tagaaaaaaa agtacttttt actccttaca attttattta
      181 cagtcaaaaa gtacttattt tttggagatc acttcattct attttccctt gctattacca
      241 aaccaattga attgcgctga tgcccagttt aatttaaata gatctggctc cggtgcccgt
      301 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      361 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      421 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      481 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      541 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctc
      601 cagtacgtga ttcttgatcc cgagctggag ccaggggcgg gccttgcgct ttaggagccc
      661 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
      721 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
      781 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaggatc
      841 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
      901 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
      961 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
     1021 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
     1081 gccctgctcc agggggctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
     1141 cacccacaca aaggaaaggg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
     1201 agtaccgggc gccgtccagg cacctcgatt agttctggag cttttggagt acgtcgtctt
     1261 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1321 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1381 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1441 tgtcgtgagg atcaaacccg ggctgcagga attcggcgcg ccacgcgtac cgatagcttc
     1501 gaattctgca gtcgacgata tcactagtct cgagcatttt ggcaaagaat tcggcatggt
     1561 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1621 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1681 gctgaccctg aagcttctat gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1741 gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1801 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1861 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1921 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1981 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     2041 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     2101 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2161 gagctaccag tccgccctgt tcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2221 ggagttcctg accgccgccg ggatcactga gggcatgaac gagctgtacc tcgagatggc
     2281 ggacgaggag aagctgccgc ccggctggga gaagcgcatg agccgcagct caggccgagt
     2341 gtactacttc aaccacatca ctaacgccag ccagtgggag cggcccagcg gcaacagcag
     2401 cagtggtggc aaaaacgggc agggggagcc tgccaggggt accagtgctg gtggtagtgc
     2461 tggtggtagt gctggtggtt ccggcagtgc tggtggtagt gctggtggta gtaccagtgc
     2521 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg
     2581 cagtgctggt ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc
     2641 tggtggtagt gctggtggta gtgctggtgg ttccggacca gatgtcccta gagctccagt
     2701 ggatccagca tggctgtcat tccaatttcc gggcggccgc atggtgagta aaggggaaga
     2761 actgtttact ggcgtggtgc caattctggt cgaactggat ggagatgtaa atgggcataa
     2821 atttagtgtg tctggagaag gggaaggaga cgctacatat gggaaactga cactgaaatt
     2881 tatttgtact actggaaaac tgcctgtgcc ttggcctaca ctcgtgacaa ccctgtcttg
     2941 gggagtgcag tgttttgccc ggtatcctga tcatatgaaa cagcatgatt ttttcaaatc
     3001 tgccatgcct gagggatatg tccaggaacg gacaattttc tttaaagatg atgggaatta
     3061 taaaacacgg gccgaagtga aatttgaagg ggatacactg gtgaatcgga ttgaactgaa
     3121 aggaattgat tttaaagaag atgggaatat tctgggccat aaactggaat ataattattt
     3181 cagcgataat gtctacatta cagccgataa acagaagaat gggattaaag ccaattttaa
     3241 aattcggcat aatattgaag atggaggggt gcagctcgcc gatcattatc agcagaatac
     3301 acctattggg gatgggcctg tgctgctgcc tgataatcat tatctgagca cacagtctgc
     3361 cctgagcaag gaccctaatg aaaagcggga ccatatggtc ctgctggaat ttctgactgc
     3421 cgccggcatt acactcggga tggatgaact gtctagacct aagaagaagc gtaaggtgta
     3481 agtcgacggt cgagtcgaga cgctagccac cggtcgttaa cattaagcta gaactatagt
     3541 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     3601 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3661 gtaaccatta taagctgcaa taaacaagtt aacaacgccg gccatcgatt aaactcaggt
     3721 tcagggggag gtgtgggagg ttttttaatt ctcgggtagg ggaggcgctt ttcccaaggc
     3781 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     3841 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     3901 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     3961 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     4021 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     4081 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     4141 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     4201 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     4261 gcctgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4321 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4381 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4441 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4501 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4561 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4621 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4681 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaac ccgggagatg
     4741 ggggaggcta actgagctct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4801 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4861 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4921 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4981 atgctgggga tgcggtgggc tctatggctt ctgaggttaa ttaagcttaa acaagaatct
     5041 ctagttttct ttcttgcttt tacttttact tccttaatac tcaagtacaa ttttaatgga
     5101 gtactttttt acttttactc aagtaagatt ctagccagat acttttactt ttaattgagt
     5161 aaaattttcc ctaagtactt gtactttcac ttgagtaaaa tttttgagta ctttttacac
     5221 ctctgctcga ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat
     5281 agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5341 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5461 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5521 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5881 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6121 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6721 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6781 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6901 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7021 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7081 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7141 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7201 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7321 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7381 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7441 catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7501 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     7561 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7621 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7681 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7741 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7801 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7861 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     7921 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     7981 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     8041 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     8101 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat a
//
