LOCUS       sequence-318435-       10488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for a Turquoise calcium sensor (GECI) that reports
            with lifetime and intensity changes - co-expresses a nuclear marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10488)
  AUTHORS   van der Linden FH, Mahlandt EK, Arts JJG, Beumer J, Puschhof J, de 
            Man SMA, Chertkova AO, Ponsioen B, Clevers H, van Buul JD, Postma M,
            Gadella TWJ Jr, Goedhart J
  TITLE     A turquoise fluorescence lifetime-based biosensor for quantitative 
            imaging of intracellular calcium.
  JOURNAL   Nat Commun. 2021 Dec 9;12(1):7159. doi: 10.1038/s41467-021-27249-w.
  PUBMED    34887382
REFERENCE   2  (bases 1 to 10488)
  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: "Nat 
            Commun."; date: "2021-12-9"; volume: "12(1):7159. doi"; pages: " 
            10.1038/s41467-021-27249-w"
COMMENT     Sequence Label: pLV-H2B-Maroon-P2A-3xnls-Tq-Ca-FLITS
FEATURES             Location/Qualifiers
     source          1..10488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    91..643
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(235..252)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     462..481
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      664..673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             670..1044
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             1887..2120
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2192..2313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2229..2248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2283..2302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2353..2488
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2508..2527)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2509..2527)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2537..2554)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2537..2553)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2568)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2695..3150
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2782..2801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2992..3013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3176..3280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3281..3349,3350..4141)
                     /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(3499..3518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4312..4900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4801..4820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5054..5071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5188..5209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5224..5229,5230..5247,5248..5254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5262..5278
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5267..5289
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5286..5302
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5286..5302
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5321..5341
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5323..5341
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5369..5595
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5596..5776
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5823..5948
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6441..6674
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6859..6903
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7165..7282
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        7305..7608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7609..7812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7763..7783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7809..7833
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             7969..8346
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      8359..8368
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9091..9147
                     /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             9166..9186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9190..9210
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9214..9234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      9247..9256
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9268..9324
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     CDS             9349..9597
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELY"
     primer_bind     complement(9661..9682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9922..9941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tttgtccaaa tgatgacagc gaagtaggtc tagaactagc ctagtgccat ttgttcagtg
       61 gcccgggaat tccgcccctc tcccccccta acgttactgg ccgaagccgc ttggaataag
      121 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
      181 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
      241 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
      301 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
      361 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
      421 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
      481 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
      541 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
      601 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggatat ggccacaacg
      661 gtcgccacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
      721 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
      781 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
      841 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
      901 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
      961 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     1021 ttcgtggccg aggagcagga ctgaatgcat gtattcaatc taagcaggct ttcactttct
     1081 cgccaactta caaggccttt ctgtgtaaac aatacctgaa cctttacccc gttgcccggc
     1141 aacggccagg tctgtgccaa gtgtttgctg acgcaacccc cactggctgg ggcttggtca
     1201 tgggccatca gcgcatgcgt ggaacctttt cggctcctct gccgatccat actgcggaac
     1261 tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cattatcggg actgataact
     1321 ctgttgtcct atcccgcaaa tatacatcgt ttccatggct gctaggctgt gctgccaact
     1381 ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatcct gcggacgacc
     1441 cttctcgggg tcgcttggga ctctctcgtc cccttctccg tctgccgttc cgaccgacca
     1501 cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg ccggaccgtg
     1561 tgcacttcgc ttcacctctg cacgtcgcat ggagaccacc gtgaacgccc accaaatatt
     1621 gcccaaggtc ttacataaga ggactcttgg actctcagca atgtcaacga ccgaccttga
     1681 ggcatacttc aaagactgtt tgtttaaaga ctgggaggag ttgggggagg agattaggtt
     1741 aaaggtcttt gtactaggag gctgtaggca taaattggtc tgcgcaccag caccatggcg
     1801 caatcactag agcggggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     1861 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     1921 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     1981 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     2041 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     2101 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     2161 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     2221 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2281 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     2341 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2401 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2461 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     2521 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2581 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2641 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     2701 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2761 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2821 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2881 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2941 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3001 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3061 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3121 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3181 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3241 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3301 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3361 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3421 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3481 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3541 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3601 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3661 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3721 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3781 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3841 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3901 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3961 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4021 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4081 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4141 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4441 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4981 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5041 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5101 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5161 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5221 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5281 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     5341 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     5401 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     5461 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     5521 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     5581 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5641 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5701 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5761 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     5821 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     5881 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     5941 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     6001 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     6061 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     6121 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     6181 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     6241 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     6301 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     6361 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     6421 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     6481 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     6541 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     6601 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     6661 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     6721 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     6781 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     6841 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     6901 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     6961 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     7021 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     7081 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     7141 gtgaacggat ctcgacggta tcggttttaa aagaaaaggg gggattgggg ggtacagtgc
     7201 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     7261 aattacaaaa attcaaaatt ttatcgataa gcttgggagt tccgcgttac ataacttacg
     7321 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7381 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7441 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7501 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     7561 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     7621 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     7681 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     7741 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     7801 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     7861 gacctccata gaagacaccg actctagcta gaggatcccc cgggctgcag gaattcggcg
     7921 cgccacgcgt accgatagct tcgaattctg cagtcgacga tcgccaccat gccagagcca
     7981 gcgaagtctg ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag
     8041 aaaggcggca agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag
     8101 gttctgaagc aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaat
     8161 tcgtttgtga acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac
     8221 aacaagcgct cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct
     8281 ggggagttgg ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc
     8341 gctaaggatc caccggtcgc caccatggtg agcaagggcg aggagctgat caaggagaac
     8401 atgcacacca agctgtacct gacaggcacc gtgaacaacc actacttcga atgcaccgcc
     8461 gaaggggagg gcaagcccta cgagggcacc cagaccaaca ggatcaaggt ggtgcgcgga
     8521 ggccccctgc cgttcgcatt cgacatcctg gccccctgct ttatgtacgg gagcaagacc
     8581 ttcattaacc atccacctga cattcccgat tattttaagc agtccttccc tgagggcttc
     8641 acatgggaga gaaccaccgt ttacgaagac gggggcacac tcactgctac acaggacacc
     8701 agcctccagg acggctgcct gatctacaac gtccaagtca gaggggagaa cttcccatcc
     8761 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccagtaccga gaccctgtac
     8821 cccgctgacg gcagcctgga aggcagactg gactgggccc tgaagctcgt gggcgggggc
     8881 cacctgcact gcagactgga aaccacatac agatccaaga aacccgcaaa gaacctcaag
     8941 atgcccggcg tctattttat tgacaggaga ctggaaagaa tcaaggaggc cgacaatgag
     9001 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
     9061 gggcacaaac ttaatgctag cggatctggc gcgacgaatt ttagtctact gaaacaagcg
     9121 ggagacgtgg aggaaaaccc tggacctctc gagatgggat cagatccaaa aaagaagaga
     9181 aaggtagacc cgaagaaaaa gcgtaaggtc gatcccaaaa agaaacggaa agtgggatca
     9241 ccggtcgcca ccatggtcga ctcttcacgt cgtaagtgga ataaggcagg tcacgcagtc
     9301 agagctatag gtcggctgag ctcacccgtg gttgtctata tcaccgccga caagcagaag
     9361 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     9421 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     9481 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     9541 gtcctactgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtaccag
     9601 ggtggcagcg gtggcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     9661 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     9721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     9781 gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt cgcccgctac
     9841 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     9901 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     9961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
    10021 aacatcctgg ggcacaagct ggagtacaac tactatagcg acaacacgcg tgaccaactg
    10081 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
    10141 gggacgataa caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca
    10201 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacattcgac
    10261 ttccctgagt tcctgacgat gatggcaaga aaaatgaatg acacagacag tgaagaggaa
    10321 attagagaag cgttccgcgt gtttgataag gacggcaatg gctacatcgg cgcagcagag
    10381 cttcgccacg tgatgacaga ccttggagag aagttaacag atgaggaggt tgatgaaatg
    10441 atcagggtag cagacatcga tggggatggt caggtaaact acgaagag
//
