LOCUS       4212_RcP9               9282 bp    DNA     circular SYN 20-MAY-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9282)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9282
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      10..748
                     /label=cloDF13
                     /note="/dnas_title=cloDF13"
                     /note="/vntifkey=33"
                     /note="color: #ffff00"
     modified_base   760..777
                     /label=User Junction
                     /note="User Junction"
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             complement(823..1905)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1867..1886
                     /label=LacI, reverse primer
                     /note="LacI, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(1906..1983)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     modified_base   2006..2020
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3; direction: RIGHT"
     terminator      2021..2065
                     /note="rrnB1 transcriptional terminator"
                     /note="color: #ffffff; direction: RIGHT"
     misc_difference 2066..2072
                     /label=cloning scar
                     /note="cloning scar"
                     /note="color: #a6acb3; direction: RIGHT"
     modified_base   2073..2086
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             complement(2088..3362)
                     /codon_start=1
                     /label=III
                     /note="/dnas_title=III"
                     /note="/vntifkey=4"
                     /note="color: #993366"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     misc_feature    2175..2220
                     /label=modified to remove internal promoter
                     /note="modified to remove internal promoter"
                     /note="color: #a6acb3"
     RBS             3364..3376
                     /label=SD8 RBS
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     modified_base   3377..3391
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3"
     promoter        3396..3423
                     /note="pspF P3"
                     /note="color: #ffffff; direction: RIGHT"
     -35_signal      3396..3401
                     /label=-35
                     /note="-35"
                     /note="color: #a6acb3; direction: RIGHT"
     -10_signal      3418..3423
                     /label=-10
                     /note="-10"
                     /note="color: #a6acb3; direction: RIGHT"
     prim_transcript complement(3430)
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    3432..3450
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     misc_feature    complement(3451)
                     /label=Transcription Start Site
                     /note="color: #a6acb3; direction: LEFT"
     promoter        3462..3478
                     /note="Sigma54 Core Promoter"
                     /note="color: #ffffff"
     -10_signal      3462..3466
                     /label=-10
                     /note="/created_by=tinawang"
                     /note="color: #a6acb3"
     -10_signal      3463..3464
                     /label=-12
                     /note="-12"
                     /note="color: #a6acb3"
     -35_signal      3471..3478
                     /label=-35
                     /note="/created_by=tinawang"
                     /note="color: #a6acb3"
     -35_signal      3475..3476
                     /label=-24
                     /note="-24"
                     /note="color: #a6acb3"
     protein_bind    3480..3512
                     /note="High Affinity IHF Site"
                     /note="color: #31849b"
     promoter        3510..3538
                     /note="pspF P1"
                     /note="color: #ffffff; direction: RIGHT"
     -35_signal      3510..3515
                     /label=-35
                     /note="-35"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        3529..3576
                     /note="Protected by pspF"
                     /note="color: #a6acb3"
     promoter        3529..3556
                     /note="pspF P2"
                     /note="color: #ffffff; direction: RIGHT"
     -35_signal      3529..3534
                     /label=-35
                     /note="-35"
                     /note="color: #a6acb3; direction: RIGHT"
     -10_signal      3533..3538
                     /label=-10
                     /note="-10"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    3540..3558
                     /note="UAS I"
                     /note="color: #31849b"
     prim_transcript complement(3544)
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    3545..3554
                     /label=pspF
                     
                     /note="/%_Identity=100"
                     /note="/Motif=TGTAATTCGC"
                     /note="/annotation_group=TGTAATTCGC: 6,240 -> 6,249"
                     /note="/modified_by=tinawang"
                     /note="color: #31849b"
     -10_signal      3551..3556
                     /label=-10
                     /note="-10"
                     /note="color: #a6acb3; direction: RIGHT"
     RBS             3560..3569
                     /label=pspF RBS
                     /note="pspF RBS"
                     /note="color: #a6acb3"
     prim_transcript complement(3560)
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    3562..3576
                     /note="UAS II"
                     /note="color: #31849b"
     protein_bind    3565..3574
                     /label=pspF
                     /note="/%_Identity=100"
                     /note="/Motif=TGAAATTCGC"
                     /note="/annotation_group=TGAAATTCGC: 6,220 -> 6,229"
                     /note="/modified_by=tinawang"
                     /note="color: #31849b"
     misc_feature    3577..3589
                     /label=USER linker
                     /note="USER linker"
                     /note="color: #a6acb3; direction: RIGHT"
     terminator      3597..4021
                     /note="rrnB1 transcriptional terminator"
                     /note="color: #ffffff"
     terminator      complement(3775..3819)
                     /note="rrnB1 transcriptional terminator"
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    3854
                     /label=X
                     /note="Removed BsaI Site"
                     /note="color: #a6acb3"
     misc_feature    join(4022..4030,4031..4033)
                     /label=USER linker
                     /note="USER linker"
                     /note="This forward directional feature has 2 segments:
                        1: 4022 .. 4030 / #a6acb3
                        2: 4031 .. 4033 / #a6acb3"
     promoter        4034..4108
                     /label=pLlacO-1
                     /label=pLlac
                     /note="Modified phage lambda PL promoter with LacI operator
                     sites (Lutz and Bujard, 1997).
                     Placed under control of Mutagenesis. (Brodel et al, 2020)
                     "
                     /note="color: #3366ff; direction: RIGHT"
     protein_bind    4038..4054
                     /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)."
                     /note="color: #31849b"
     protein_bind    4060..4076
                     /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)."
                     /note="color: #31849b"
     gene            4185..4997
                     /standard_name="PF020"
                     /label=PF020
                     /note="color: #a6acb3; direction: RIGHT"
     gene            5016..6863
                     /standard_name="mutL E32K"
                     /label=mutL E32K
                     /note="color: #a6acb3; direction: RIGHT"
     RBS             6864..6876
                     /label=SD8 RBS
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     gene            6877..7839
                     /label=ec86RT
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    7855..7945
                     /label=msr
                     /note="color: #85dae9"
     misc_feature    7855..7864
                     /label=extended a1
                     /note="color: #ff6600"
     misc_feature    7865..7876
                     /label=inverted repeat
                     /label=nonstandard type:  IR
                     /note="color: #b1ff67; direction: RIGHT"
     misc_feature    7935..7963
                     /label=msd
                     /note="color: #99ccff"
     misc_feature    7935..7951
                     /label=Flanking
                     /note="color: #00ff00"
     misc_feature    7952..7963
                     /label=stem base
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    7971..8345
                     /label=ccdB Selection
                     /note="Insertion of ccdB Gene for counter selection against
                     uncut DNA during oligo insertion"
                     /note="color: #ff9900"
     misc_feature    7971..8034
                     /label=Promotor Region
                     /note="color: #a6acb3"
     misc_feature    7971..7975
                     /label=-24 Region
                     /note="color: #a6acb3"
     misc_feature    7982..7986
                     /label=-12 Region
                     /note="color: #a6acb3"
     misc_feature    8023..8034
                     /label=MCS
                     /note="color: #a6acb3"
     CDS             8040..8345
                     /codon_start=1
                     /label=ccdB
                     /note="color: #000000"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    8353..8406
                     /label=msd
                     /note="color: #99ccff"
     misc_feature    complement(8357..8368)
                     /label=stem base
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    complement(8369..8383)
                     /label=Flanking
                     /note="/created_by=Seth"
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    complement(8383)
                     /label=RT Start
                     /note="color: #ffcc00; direction: LEFT"
     misc_feature    complement(8385..8396)
                     /label=inverted repeat
                     /label=nonstandard type:  IR
                     /note="color: #b1ff67; direction: LEFT"
     misc_feature    8397..8406
                     /label=extended a2
                     /note="/created_by=User"
                     /note="/modified_by=Seth"
                     /note="color: #ff6600"
     misc_feature    8407..8423
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3; direction: RIGHT"
     terminator      8431..8466
                     /note="P14/tonB bidirectional terminator"
                     /note="termination of cat transcript is slightly weakerthan
                     in opposite direction"
                     /note="color: #ffffff"
     modified_base   8467..8479
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             complement(8480..9139)
                     /label=cat (CmR)
                     /note="cat (CmR)"
                     /note="from pACYCDuet-1"
                     /note="color: #993366; direction: LEFT"
     modified_base   8555
                     /label=mutation
                     /note="mutation"
                     /note="annotated as a G in pACYCDuet cat markerannotation, 
                     here it is an A, but this mutation is silent from codon GTC
                     (Val, 25% codon usage) to GTT (Val, 21% usage), so it 
                     should not be of functional relevance"
                     /note="color: #a6acb3"
     misc_feature    8705
                     /label=C to A
                     /note="Removed Esp3I Site"
                     /note="color: #a6acb3"
     promoter        9140..9237
                     /note="cat promoter"
                     /note="from pACYCDuet-1"
                     /note="color: #ffffff; direction: RIGHT"
     -35_signal      9232..9239
                     /label=-35
                     /note="/created_by=tinawang"
                     /note="color: #a6acb3"
     modified_base   9238..9253
                     /label=USER junction
                     /note="USER junction"
                     /note="color: #a6acb3; direction: RIGHT"
ORIGIN
        1 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
       61 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
      121 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
      181 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
      241 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
      301 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
      361 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
      421 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
      481 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
      541 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
      601 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
      661 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
      721 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca
      781 atttatctct tcaaatgtag cacctgaagt cagcccagga ggtcactgcc cgctttccag
      841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      901 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gacacgggca acagctgatt
      961 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     1021 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     1081 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     1141 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatacc
     1201 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     1261 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     1321 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     1381 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     1441 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     1501 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     1561 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     1621 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     1681 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     1741 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     1801 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     1861 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     1921 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     1981 gtccgggatc tcgacgctct cccttaagag gacatccggt caaataaaac gaaaggctca
     2041 gtcgaaagac tgggcctttc gttttgctga ggagacttag ggacccttta agactcctta
     2101 ttacgcagta tgttagcaaa cgtagaaaat acatacataa aggtggcaac atataaaaga
     2161 aacgcaaaga caccgcggaa caggttgatc ttatcgcagt cgatactgaa ctcgtaaggt
     2221 ttaccagcgc caaagacaaa agggcgacat tcaaccgatt gagggaggga aggtaaatat
     2281 tgacggaaat tattcattaa aggtgaatta tcaccgtcac cgacttgagc catttgggaa
     2341 ttagagccag caaaatcacc agtagcacca ttaccattag caaggccgga aacgtcacca
     2401 atgaaaccat cgatagcagc accgtaatca gtagcgacag aatcaagttt gcctttagcg
     2461 tcagactgta gcgcgttttc atcggcattt tcggtcatag cccccttatt agcgtttgcc
     2521 atcttttcat aatcaaaatc accggaacca gagccaccac cggaaccgcc tccctcagag
     2581 ccgccaccct cagaaccgcc accctcagag ccaccaccct cagagccgcc accagaacca
     2641 ccaccagagc cgccgccagc attgacagga ggttgaggca ggtcagacga ttggccttga
     2701 tattcacaaa cgaatggatc ctcattaaag ccagaatgga aagcgcagtc tctgaattta
     2761 ccgttccagt aagcgtcata catggctttt gatgatacag gagtgtactg gtaataagtt
     2821 ttaacggggt cagtgccttg agtaacagtg cccgtataaa cagttaatgc cccctgccta
     2881 tttcggaacc tattattctg aaacatgaaa gtattaagag gctgagactc ctcaagagaa
     2941 ggattaggat tagcggggtt ttgctcagta ccaggcggat aagtgccgtc gagagggttg
     3001 atataagtat agcccggaat aggtgtatca ccgtactcag gaggtttagt accgccaccc
     3061 tcagaaccgc caccctcaga accgccaccc tcagagccac caccctcatt ttcagggata
     3121 gcaagcccaa taggaaccca tgtaccgtaa cactgagttt cgtcaccagt acaaactaca
     3181 acgcctgtag cattccacag acagccctca tagttagcgt aacgatctaa agttttgtcg
     3241 tctttccaga cgttagtaaa tgaattttct gtatggggtt ttgctaaaca actttcaaca
     3301 gtttcagcgg agtgagaata gaaaggaaca actaaaggaa ttgcgaataa taattttttc
     3361 attttttttt tcctttactg cacctgcagg taatgttgtc ctcttgattt ctgcgttcag
     3421 gattgtcctg ctctctatca ctgataggga tgaactgtta atacaatttg cgtgccaatt
     3481 ttttatcttt ttgatttata aagatctgat tgaagaatca acagcaacat gccaggatga
     3541 gttagcgaat tacactaaca agtggcgaat ttcatcacgg agccaatgtc ctcagcgagt
     3601 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc
     3661 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc
     3721 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3781 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3841 tcgcatgggg agaacccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     3901 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc
     3961 ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc
     4021 cagggcccta ctgataaatg tgagcggata acattgacat tgtgagcgga taacaagata
     4081 ctgagcacat cagcaggacg cactgacctc aacagaaaca ataataatgg agtcatgacc
     4141 atgcctaggc cgcgggccgc gcgaattcag aaggagaata taccatgaac caaatcgtga
     4201 agttcactga cgactctggc ctggcggttc aagttactcc agacgatgtt cgccgttata
     4261 tctgtgagaa cgctactgaa aaagaggtgg gcctctttct gcaactctgt cagactcaac
     4321 gtctgaatcc gtttgtgaaa gacgcttacc tggtgaaata cggcggtgct ccagcttcta
     4381 tgattacttc ctatcaagtt tttaaccgtc gcgcgtgtcg tgatgctaac tatgatggta
     4441 tcaaatctgg tgtggttgtt ctgcgtgacg gtgatgttgt gcataaacgt ggtgctgcgt
     4501 gctacaaaaa ggcgggtgag gagctcatcg gtggttgggc ggaagttcgc tttaaggatg
     4561 gccgcgagac tgcgtatgct gaggtggcgc tcgacgacta ttccaccggc aaatctaatt
     4621 gggcgaaaat gccgggtgtt atgatcgaaa aatgcgcgaa ggctgctgct tggcgcctcg
     4681 cgttcccgga cacttttcag ggcatgtacg ctgcggagga aatggatcaa gcgcaacagc
     4741 cagaacaggt gcgcgctcag gcggagcaac cagtggatct ccagccaatc cgcgaactct
     4801 tcaagccata ttgcgaacac ttcggcatca ctccggctga gggtatgact gctgtttgtg
     4861 gtgcggtggg cgctgaaggc atgcactcta tgaccgagca gcaagctcgc cgtgctcgcg
     4921 cttggatgga ggaagaaatg gctgcgccag ctgtggaagc ggagtatgag gttgttgacg
     4981 agggcgaggt gttttaagga tccaggagaa tacacatgcc aattcaggtc ttaccgccac
     5041 aactggcgaa ccagattgcc gcaggtgagg tggtcgagcg acctgcgtcg gtagtcaaag
     5101 aactagtgaa aaacagcctc gatgcaggtg cgacgcgtat cgatattgat atcgaacgcg
     5161 gtggggcgaa acttatccgc attcgtgata acggctgcgg tatcaaaaaa gatgagctgg
     5221 cgctggcgct ggctcgtcat gccaccagta aaatcgcctc tctggacgat ctcgaagcca
     5281 ttatcagcct gggctttcgc ggtgaggcgc tggcgagtat cagttcggtt tcccgcctga
     5341 cgctcacttc acgcaccgca gaacagcagg aagcctggca ggcctatgcc gaagggcgcg
     5401 atatgaacgt gacggtaaaa ccggcggcgc atcctgtggg gacgacgctg gaggtgctgg
     5461 atctgttcta caacaccccg gcgcggcgca aattcctgcg caccgagaaa accgaattta
     5521 accacattga tgagatcatc cgccgcattg cgctggcgcg tttcgacgtc acgatcaacc
     5581 tgtcgcataa cggtaaaatt gtgcgtcagt accgcgcagt gccggaaggc gggcaaaaag
     5641 aacggcgctt aggcgcgatt tgcggcaccg cttttcttga acaagcgctg gcgattgaat
     5701 ggcaacacgg cgatctcacg ctacgcggct gggtggccga tccaaatcac accacgcccg
     5761 cactggcaga aattcagtat tgctacgtga acggtcgcat gatgcgcgat cgcctgatca
     5821 atcacgcgat ccgccaggcc tgcgaagaca aactgggggc cgatcagcaa ccggcatttg
     5881 tgttgtatct ggagatcgac ccacatcagg tggacgtcaa cgtgcacccc gccaaacacg
     5941 aagtgcgttt ccatcagtcg cgtctggtgc atgattttat ctatcagggc gtgctgagcg
     6001 tgctacaaca gcaactggaa acgccgctac cgctggacga tgaaccccaa cctgcaccgc
     6061 gttccattcc ggaaaaccgc gtggcggcgg ggcgcaatca ctttgcagaa ccggcagctc
     6121 gtgagccggt agctccgcgc tacactcctg cgccagcatc aggcagtcgt ccggctgccc
     6181 cctggccgaa tgcgcagcca ggctaccaga aacagcaagg tgaagtgtat cgccagcttt
     6241 tgcaaacgcc cgcgccgatg caaaaattaa aagcgccgga accgcaggaa cctgcacttg
     6301 cggcgaacag tcagagtttt ggtcgggtac tgactatcgt ccattccgac tgtgcgttgc
     6361 tggagcgcga cggcaacatt tcacttttat ccttgccagt ggcagaacgt tggctgcgtc
     6421 aggcacaatt gacgccgggt gaagcgcccg tttgcgccca gccgctgctg attccgttgc
     6481 ggctaaaagt ttctgccgaa gaaaaatcgg cattagaaaa agcgcagtct gccctggcgg
     6541 aattgggtat tgatttccag tcagatgcac agcatgtgac catcagggca gtgcctttac
     6601 ccttacgcca acaaaattta caaatcttga ttcctgaact gataggctac ctggcgaagc
     6661 agtccgtatt cgaacctggc aatattgcgc agtggattgc acgaaatctg atgagcgaac
     6721 atgcgcagtg gtcaatggca caggccataa ccctgctggc ggacgtggaa cggttatgtc
     6781 cgcaacttgt gaaaacgccg ccgggtggtc tgttacaatc tgttgattta catccggcga
     6841 taaaagccct gaaagatgag tgaaaaggaa aaaaaaatga aatccgctga atatttgaac
     6901 acttttagat tgagaaatct cggcctacct gtcatgaaca atttgcatga catgtctaag
     6961 gcgactcgca tatctgttga aacacttcgg ttgttaatct atacagctga ttttcgctat
     7021 aggatctaca ctgtagaaaa gaaaggccca gagaagagaa tgagaaccat ttaccaacct
     7081 tctcgagaac ttaaagcctt acaaggatgg gttctacgta acattttaga taaactgtcg
     7141 tcatctcctt tttctattgg atttgaaaag caccaatcta ttttgaataa tgctaccccg
     7201 catattgggg caaactttat actgaatatt gatttggagg attttttccc aagtttaact
     7261 gctaacaaag tttttggagt gttccattct cttggttata atcgactaat atcttcagtt
     7321 ttgacaaaaa tatgttgtta taaaaatctg ctaccacaag gtgctccatc atcacctaaa
     7381 ttagctaatc taatatgttc taaacttgat tatcgtattc agggttatgc aggtagtcgg
     7441 ggcttgatat atacgagata tgccgatgat ctcaccttat ctgcacagtc tatgaaaaag
     7501 gttgttaaag cacgtgattt tttattttct ataatcccaa gtgaaggatt ggttattaac
     7561 tcaaaaaaaa cttgtattag tgggcctcgt agtcagagga aagttacagg tttagttatt
     7621 tcacaagaga aagttgggat aggtagagaa aaatataaag aaattagagc aaagatacat
     7681 catatatttt gcggtaagtc ttctgagata gaacacgtta ggggatggtt gtcatttatt
     7741 ttaagtgtgg attcaaaaag ccataggaga ttaataactt atattagcaa attagaaaaa
     7801 aaatatggaa agaacccttt aaataaagcg aagacctaac atttaactat gataagattc
     7861 cgtatgcgca cccttagcga gaggtttatc attaaggtca acctctggat gttgtttcgg
     7921 catcctgcat tgaatctgag ttactgtctg ttttcctgaa ttctgagacg tggcacaccc
     7981 cttgctttgt ctttatcaac gcaaataaca agttgataac aagctagcag gaggaattca
     8041 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     8101 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     8161 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     8221 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     8281 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     8341 tataacgtct cagtacgaat tcaggaaacc cgttttttct gacgtaaggg tgcgcatacg
     8401 gaatctactt aattaacggc actcctcagc caagtcaaaa gcctccgacc ggaggctttt
     8461 gactacatgc ccatggcgtt tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     8521 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     8581 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga
     8641 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     8701 ctgaaacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     8761 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     8821 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     8881 tatcccatat caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca
     8941 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     9001 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat ttagcaactg
     9061 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     9121 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     9181 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccaag
     9241 ccaaataggc cgtcactcgg tcgctacgct ccgggcgtga ga
//
