LOCUS       pJRD235-pET-Cre-        6301 bp DNA     circular SYN 18-FEB-2025
DEFINITION  Expression of Cre-LPETG-6xHis in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6301)
  AUTHORS   Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder
            ML, Joung JK, Chen ZY, Liu DR
  TITLE     Cationic lipid-mediated delivery of proteins enables efficient 
            protein-based genome editing in vitro and in vivo.
  JOURNAL   Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 
            2014 Oct 30.
  PUBMED    25357182
REFERENCE   2  (bases 1 to 6301)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2025 from SnapGene 7.2.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt.3081"; 
            journalName: "Nat Biotechnol"; date: "2015-01"; volume: "33"; issue:
            "1"; pages: "73-80"
FEATURES             Location/Qualifiers
     source          1..6301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1094..1198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1199..1228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(44..73)
                     /label=JRD149
                     /note="/%GC=30.0"
                     /note="/Sequence=CATGGTATATCTCCTTCTTAAAGTTAAACA"
                     /note="/Transferred_From=JRD149"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(51..70)
                     /label=JD116
                     /note="/%GC=35.0"
                     /note="/Sequence=GGTATATCTCCTTCTTAAAG"
                     /note="/Transferred_From=JD116"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     54..93
                     /label=JRD740
                     /note="/Sequence=TAAGAAGGAGATATACCATGAATTTACTGACCGTACACCA"
                     /note="/created_by=jessiedavis"
                     /note="/modified_by=jessiedavis"
     CDS             71..1093
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     G"
     primer_bind     complement(195..214)
                     /label=CRE-R
                     /label=CRE-R; QB0013
                     /note="/%GC=45.0"
                     /note="/Sequence=GCAAACGGACAGAAGCATTT"
                     /note="/Transferred_From=CRE-R, QB0013"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     338..357
                     /label=JRD357
                     /note="/%GC=45.0"
                     /note="/Sequence=CATTTGGGCCAGCTAAACAT"
                     /note="/Transferred_From=JRD357"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(552..574)
                     /label=JRD358
                     /note="/%GC=39.1"
                     /note="/Sequence=GTTATAAGCAATCCCCAGAAATG"
                     /note="/Transferred_From=JRD358"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(561..584)
                     /label=JRD359
                     /note="/%GC=41.7"
                     /note="/Sequence=GTAACAGGGTGTTATAAGCAATCC"
                     /note="/Transferred_From=JRD359"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     1074..1106
                     /label=JRD742
                     /note="/%GC=66.7"
                     /note="/Hairpin_Tm=48.5"
                     /note="/Self_Dimer_Tm=34.6"
                     /note="/Sequence=TGCGCCTGCTGGAAGATGGCACCGGTGGTAGCG"
                     /note="/Tm=78.5"
                     /note="/created_by=primer3"
                     /note="/modified_by=jessiedavis"
     primer_bind     complement(1079..1148)
                     /label=JRD746
                     /note="/Sequence=TACCACCGCTACCGCCGCTACCACCAGAACCACCAGAACCAC
                     CGCTA CCACCGGTGCCATCTTCCAGCAG"
                     /note="/created_by=jessiedavis"
                     /note="/modified_by=jessiedavis"
     primer_bind     complement(1079..1093)
                     /label=JRD741
                     /note="/%GC=60.0"
                     /note="/Hairpin_Tm=None"
                     /note="/Self_Dimer_Tm=None"
                     /note="/Sequence=GCCATCTTCCAGCAG"
                     /note="/Tm=50.5"
                     /note="/created_by=primer3"
     misc_feature    1094..1198
                     /label=Untitled
                     /label=nonstandard type: Editing History Insertion
                     /note="/Original_Bases"
     misc_feature    1094..1177
                     /label=Linker
                     /note="/created_by=jessiedavis"
     primer_bind     1129..1191
                     /label=JRD745
                     /note="/Sequence=TAGCGGCGGTAGCGGTGGTAGCGGTGGTAGCGGTGGCAGCGG
                     CGGTA CCCTCCCAGAGACCGG"
                     /note="/created_by=jessiedavis"
                     /note="/modified_by=jessiedavis"
     misc_feature    1178..1198
                     /label=Sortag
                     /note="/created_by=jessiedavis"
     primer_bind     complement(1183..1213)
                     /label=JRD747
                     /note="/%GC=61.3"
                     /note="/Hairpin_Tm=42.4"
                     /note="/Self_Dimer_Tm=2.6"
                     /note="/Sequence=ATGGGATCCGCCGTCTACGACACCGGTCTCT"
                     /note="/Tm=74.2"
                     /note="/created_by=primer3"
     primer_bind     complement(1183..1198)
                     /label=JRD743
                     /note="/%GC=56.3"
                     /note="/Hairpin_Tm=42.1"
                     /note="/Self_Dimer_Tm=2.6"
                     /note="/Sequence=TACGACACCGGTCTCT"
                     /note="/Tm=53.0"
                     /note="/created_by=primer3"
                     /note="/modified_by=jessiedavis"
     misc_feature    join(1198,1229)
                     /label=junction 1
                     /label=nonstandard type: Junction
                     /note="/Deletion_Size=30"
                     /note="/ID=junction 1"
                     /note="/Junction_Destination=1229+"
                     /note="/Junction_Source=1198+"
                     /note="/Junction_Type=Deletion"
                     /note="/Reads_supporting_discovery=4"
                     /note="/Reads_using=4"
     primer_bind     1207..1246
                     /label=JRD748
                     /note="/Sequence=ATCCCATCACCACCACCATCACTAATGACTCGAGTCTGGT"
                     /note="/created_by=jessiedavis"
                     /note="/modified_by=jessiedavis"
     misc_feature    1207..1213
                     /label=User jxn
                     /label=nonstandard type: USER
                     /note="/created_by=jessiedavis"
     CDS             1211..1228
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1247..1291
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1305..1327)
                     /label=HR166
                     /note="/%GC=47.8"
                     /note="/Sequence=GGCAGCAGCCTAGGTTAATTAAG"
                     /note="/Transferred_From=HR166"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(1329..1347)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1343..1390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1427..1882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1453..1485
                     /label=NMG-306
                     /note="/%GC=72.7"
                     /note="/Sequence=GCGGCGGGTGTGGTGGTTACGCGCAGCGTGACC"
                     /note="/Transferred_From=NMG-306"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(1514..1533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1724..1745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1989..2017
                     /label=NMG-853
                     /note="/%GC=37.9"
                     /note="/Sequence=TCTGACAGTTACCAATGCTTAATCAGTGA"
                     /note="/Transferred_From=NMG-853"
                     /note="/Transferred_Similarity=100.00%"
     CDS             complement(1997..2857)
                     /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     2147..2176
                     /label=NMG-584
                     /note="/%GC=43.3"
                     /note="/Sequence=CTCACCGGCTCCAGATTTATCAGCAATAAA"
                     /note="/Transferred_From=NMG-584"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2238..2262
                     /label=ACK-12
                     /note="/%GC=40.0"
                     /note="/Sequence=TCTATTAATTGTTGCCGGGAAGCTA"
                     /note="/Transferred_From=ACK-12"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2346..2372
                     /label=NMG-1102
                     /note="/%GC=51.9"
                     /note="/Sequence=ATGGCTTCATTCAGCTCCGGTTCCCAA"
                     /note="/Transferred_From=NMG-1102"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2521..2549
                     /label=NMG-854
                     /note="/%GC=48.3"
                     /note="/Sequence=CCGTAAGATGCTTTTCTGTGACTGGTGAG"
                     /note="/Transferred_From=NMG-854"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2620..2639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2681..2710
                     /label=NMG-585
                     /note="/%GC=46.7"
                     /note="/Sequence=GCGAAAACTCTCAAGGATCTTACCGCTGTT"
                     /note="/Transferred_From=NMG-585"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(2724..2751)
                     /label=JRD750
                     /note="/%GC=50.0"
                     /note="/Hairpin_Tm=None"
                     /note="/Self_Dimer_Tm=11.5"
                     /note="/Sequence=ATCAGTTGGGTGCACGAGTGGGTTACAT"
                     /note="/Tm=68.1"
                     /note="/created_by=primer3"
                     /note="/modified_by=jessiedavis"
     primer_bind     2738..2762
                     /label=ACK-13
                     /note="/%GC=48.0"
                     /note="/Sequence=TGCACCCAACTGATCTTCAGCATCT"
                     /note="/Transferred_From=ACK-13"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2741..2771
                     /label=JRD749
                     /note="/%GC=38.7"
                     /note="/Hairpin_Tm=48.1"
                     /note="/Self_Dimer_Tm=4.5"
                     /note="/Sequence=ACCCAACTGATCTTCAGCATCTTTTACTTTC"
                     /note="/Tm=63.9"
                     /note="/created_by=primer3"
     misc_feature    2741..2751
                     /label=nonstandard type: USER
                     /note="/created_by=jessiedavis"
     primer_bind     2839..2868
                     /label=NMG-396
                     /note="/%GC=33.3"
                     /note="/Sequence=GGAAATGTTGAATACTCATACTCTTCCTTT"
                     /note="/Transferred_From=NMG-396"
                     /note="/Transferred_Similarity=100.00%"
     promoter        complement(2858..2950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2897..2919
                     /label=NMG-1469
                     /note="/%GC=43.5"
                     /note="/Sequence=GTTATTGTCTCATGAGCGGATAC"
                     /note="/Transferred_From=NMG-1469"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     2913..2939
                     /label=NMG-1103
                     /note="/%GC=25.9"
                     /note="/Sequence=CGGATACATATTTGAATGTATTTAGAA"
                     /note="/Transferred_From=NMG-1103"
                     /note="/Transferred_Similarity=100.00%"
     rep_origin      3031..3619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3094..3118)
                     /label=ACK-11
                     /note="/%GC=52.0"
                     /note="/Sequence=CTCTTGATCCGGCAAACAAACCACC"
                     /note="/Transferred_From=ACK-11"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     3252..3271
                     /label=QB0767
                     /label=QB0767; RepO-R
                     /note="/%GC=55.0"
                     /note="/Sequence=TCCTGTTACCAGTGGCTGCT"
                     /note="/Transferred_From=QB0767, RepO-R"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     3295..3321
                     /label=NMG-309
                     /note="/%GC=48.1"
                     /note="/Sequence=TACCGGGTTGGACTCAAGACGATAGTT"
                     /note="/Transferred_From=NMG-309"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(3378..3396)
                     /label=QB0766
                     /label=QB0766; RepO-F
                     /note="/%GC=57.9"
                     /note="/Sequence=GTGTAGGTCGTTCGCTCCA"
                     /note="/Transferred_From=QB0766, RepO-F"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(3384..3411)
                     /label=NMG-851
                     /note="/%GC=50.0"
                     /note="/Sequence=AGGTATCTCAGTTCGGTGTAGGTCGTTC"
                     /note="/Transferred_From=NMG-851"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     3520..3539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3595..3619)
                     /label=ACK-10
                     /note="/%GC=64.0"
                     /note="/Sequence=TTTCCATAGGCTCCGCCCCCCTGAC"
                     /note="/Transferred_From=ACK-10"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     3773..3790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3805..3947
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3837..3869
                     /label=NMG-310
                     /note="/%GC=48.5"
                     /note="/Sequence=GCGGTATTTCACACCGCATATATGGTGCACTCT"
                     /note="/Transferred_From=NMG-310"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(3873..3892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4033..4055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4049..4240)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low
                     copy number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4373..4402
                     /label=NMG-311
                     /note="/%GC=63.3"
                     /note="/Sequence=GGCGGTATGGATGCGGCGGGACCAGAGAAA"
                     /note="/Transferred_From=NMG-311"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     complement(4770..4789)
                     /label=Lac F
                     /label=Lac F; Lac-F
                     /note="/%GC=50.0"
                     /note="/Sequence=TAGCTCACTCATTAGGCACC"
                     /note="/Transferred_From=Lac F, Lac-F"
                     /note="/Transferred_Similarity=100.00%"
     CDS             complement(4812..5894)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4836..4862
                     /label=NMG-1101
                     /note="/%GC=48.1"
                     /note="/Sequence=AACCTGTCGTGCCAGCTGCATTAATGA"
                     /note="/Transferred_From=NMG-1101"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     5202..5229
                     /label=NMG-313
                     /note="/%GC=42.9"
                     /note="/Sequence=TTGCATGGTTTGTTGAAAACCGGACATG"
                     /note="/Transferred_From=NMG-313"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     5724..5753
                     /label=NMG-314
                     /note="/%GC=60.0"
                     /note="/Sequence=CGCCAGTTGTTGTGCCACGCGGTTGGGAAT"
                     /note="/Transferred_From=NMG-314"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     5856..5875
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5895..5972)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6012..6031
                     /label=JRDSeq2
                     /note="/%GC=60.0"
                     /note="/Sequence=GGAAGCAGCCCAGTAGTAGG"
                     /note="/Transferred_From=JRDSeq2"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     6178..6197
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     primer_bind     6224..6245
                     /label=HR129
                     /note="/%GC=68.2"
                     /note="/Sequence=GTGATGCCGGCCACGATGCGTC"
                     /note="/Transferred_From=HR129"
                     /note="/Transferred_Similarity=100.00%"
     primer_bind     join(6285..6301,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6285..6301,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(6285..6301,1..2)
                     /note="/%GC=36.8"
                     /note="/Sequence=TAATACGACTCACTATAGG"
                     /note="/Transferred_From"
                     /note="/Transferred_Similarity=100.00%"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgaatttac tgaccgtaca ccaaaatttg cctgcattgc cggtcgatgc
      121 aacgagtgat gaggttcgca agaacctgat ggacatgttc agggatcgcc aggcgttttc
      181 tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt
      241 gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc ttctatatct
      301 tcaggcgcgc ggtctggcag taaaaactat ccagcaacat ttgggccagc taaacatgct
      361 tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat gctgtttcac tggttatgcg
      421 gcgtatccga aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga
      481 acgcactgat ttcgaccagg ttcgttcact catggaaaat agcgatcgct gccaggatat
      541 acgtaatctg gcatttctgg ggattgctta taacaccctg ttacgtatag ccgaaattgc
      601 caggatcagg gttaaagata tctcacgtac tgacggtggg agaatgttaa tccatattgg
      661 cagaacgaaa acgctggtta gcaccgcagg tgtagagaag gcacttagcc tgggggtaac
      721 taaactggtc gagcgatgga tttccgtctc tggtgtagct gatgatccga ataactacct
      781 gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc agctatcaac
      841 tcgcgccctg gaagggattt ttgaagcaac tcatcgattg atttacggcg ctaaggatga
      901 ctctggtcag agatacctgg cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga
      961 tatggcccgc gctggagttt caataccgga gatcatgcaa gctggtggct ggaccaatgt
     1021 aaatattgtc atgaactata tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct
     1081 gctggaagat ggcaccggtg gtagcggtgg ttctggtggt tctggtggta gcggcggtag
     1141 cggtggtagc ggtggtagcg gtggcagcgg cggtaccctc ccagagaccg gtgtcgtaga
     1201 cggcggatcc catcaccacc accatcacta atgactcgag tctggtaaag aaaccgctgc
     1261 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc
     1321 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1381 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     1441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1501 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1561 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1621 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1681 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1741 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1801 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1861 aaaatattaa cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct
     1921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat
     2881 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2941 aataaacaaa taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3001 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3061 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3781 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3841 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3901 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3961 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4021 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4081 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4141 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4201 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4261 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4321 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4381 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4441 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4501 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     4561 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     4621 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     4681 acaggagcac gatcatgcta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     4741 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     4801 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4861 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     4921 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     4981 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     5041 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     5101 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     5161 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     5221 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     5281 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     5341 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     5401 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     5461 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     5521 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     5581 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     5641 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     5701 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     5761 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     5821 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     5881 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     5941 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     6001 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     6061 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     6121 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     6181 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     6241 gcgtccggcg tagaggatcg agatcgatct cgatcccgcg aaattaatac gactcactat
     6301 a
//
