LOCUS       sequence-433660-       14636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduction of low-temperature-inducible GAL system, LowTempGAL 
            v2.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14636)
  AUTHORS   Lu Z, Shen Q, Bandari NC, Evans S, McDonnell L, Liu L, Jin W, 
            Luna-Flores CH, Collier T, Talbo G, McCubbin T, Esquirol L, Myers C,
            Trau M, Dumsday G, Speight R, Howard CB, Vickers CE, Peng B
  TITLE     LowTempGAL: a highly responsive low temperature-inducible GAL system
            in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2024 May 29:gkae460. doi: 10.1093/nar/gkae460.
  PUBMED    38808673
REFERENCE   2  (bases 1 to 14636)
  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: "Nucleic 
            Acids Res. 2024 May 29:gkae460. doi: 10.1093/nar/gkae460."
COMMENT     Sequence Label: pIRHD-TetR-GAL80-Rho1-GAL4
FEATURES             Location/Qualifiers
     source          1..14636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1313..3955
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDFM
                     PRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVNR
                     LPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQIL
                     FNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQK
                     TNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRSI
                     QLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICAK
                     KCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVLR
                     DFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAWN
                     CSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYIQ
                     VLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKYV
                     SPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQLQ
                     SLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSNV
                     HDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDDE
                     DTPPNPKKE"
     protein_bind    4879..4897
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4906..4924
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4940..4958
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4966..4984
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(5931..5947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5931..5947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5944..5966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5955..5971
                     /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(join(5979..5985,5986..6003,6004..6009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6024..6045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6162..6179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6333..6921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6413..6432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7092..7883,7884..7952))
                     /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     7715..7734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7953..8057)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8125..8143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8181..8203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8303..8322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8516..8538
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8530..8547
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8531..8547
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            8889..10245
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        8889..9232
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             9233..10042
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(9300..9319)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      10048..10245
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             10844..11071
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
ORIGIN
        1 gtattaatca catgtatatc gcggccgcac tttgttatgt agagtttttt tagctaccta
       61 tattccacca taacatcaat catgcggttg ctggtgtatt taccaataat gtttaatgta
      121 tatatatata tatatatggg gccgtatact tacatatagt agatgtcaag cgtaggcgct
      181 tcccctgccg gctgtgaggg cgccataacc aaggtatcta tagaccgcca atcagcaaac
      241 tacctccgta cattcatgtt gcacccacac atttatacac ccagaccgcg acaaattacc
      301 cataaggttg tttgtgacgg cgtcgtacaa gagaacgtgg gaacttttta ggctcaccaa
      361 aaaagaaaga aaaaatacga gttgctgaca gaagcctcaa gaaaaaaaaa attcttcttc
      421 gactatgctg gaggcagaga tgatcgagcc ggtagttaac tatatatagc taaattggtt
      481 ccatcacctt cttttctggt gtcgctcctt ctagtgctat ttctggcttt tcctattttt
      541 ttttttccat ttttctttct ctctttctaa tatataaatt ctcttgcatt ttctattttt
      601 ctctctatct attctacttg tttattccct tcaaggtttt tttttaagga gtacttgttt
      661 ttagaatata cggtcaacga actataatta actaaacatg gctaccgaat tgagaagaaa
      721 gttggttata gttggtgatg gtgctagagg taagacttgt ttgttgatcg ttttctccaa
      781 gggtactttc ccagaagttt acgttccaac cgtttttgaa aactacgttg ccgatgttga
      841 agttgatggt agacatgttg aattggcttt gtgggatact gctggtcaag aagattacga
      901 tagactaaga ccattgtctt acccagattc tcacgttatt ttgatttgct tcgctgttga
      961 ttctccagac tctttggata acgttcaaga aaagtggatc tccgaagtct tgcatttctg
     1021 ttcttcattg ccaattttgt tggttgcttg caaggctgat ttgagaaacg atccaaagat
     1081 catcgaagag ttgtccaaga ctaatcaaca tccagttact accgaagaag gtcaagctgt
     1141 tgctcaaaaa attggtgcct acaaatactt ggaatgctct gctaaaacta acgaaggtgt
     1201 cagagaagtt ttcgaatctg ctactagagc tgctatgttg aaacataagc caaaggttaa
     1261 gccatcctcc ggtactaaga aaaagaagag atgcattcta ctaggttctg gtaagctact
     1321 gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga
     1381 aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac
     1441 caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact
     1501 ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga
     1561 ttctttacag gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa
     1621 agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca
     1681 gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt
     1741 gactgtatcg attgactcgg cagctcatca tgataactcc acaattccgt tggattttat
     1801 gcccagggat gctcttcatg gatttgattg gtctgaagag gatgacatgt cggatggctt
     1861 gcccttcctg aaaacggacc ccaacaataa tgggttcttt ggcgacggtt ctctcttatg
     1921 tattcttcga tctattggct ttaaaccgga aaattacacg aactctaacg ttaacaggct
     1981 cccgaccatg attacggata gatacacgtt ggcttctaga tccacaacat cccgtttact
     2041 tcaaagttat ctcaataatt ttcaccccta ctgccctatc gtgcactcac cgacgctaat
     2101 gatgttgtat aataaccaga ttgaaatcgc gtcgaaggat caatggcaaa tcctttttaa
     2161 ctgcatatta gccattggag cctggtgtat agagggggaa tctactgata tagatgtttt
     2221 ttactatcaa aatgctaaat ctcatttgac gagcaaggtc ttcgagtcag gttccataat
     2281 tttggtgaca gccctacatc ttctgtcgcg atatacacag tggaggcaga aaacaaatac
     2341 tagctataat tttcacagct tttccataag aatggccata tcattgggct tgaataggga
     2401 cctcccctcg tccttcagtg atagcagcat tctggaacaa agacgccgaa tttggtggtc
     2461 tgtctactct tgggagatcc aattgtccct gctttatggt cgatccatcc agctttctca
     2521 gaatacaatc tccttccctt cttctgtcga cgatgtgcag cgtaccacaa caggtcccac
     2581 catatatcat ggcatcattg aaacagcaag gctcttacaa gttttcacaa aaatctatga
     2641 actagacaaa acagtaactg cagaaaaaag tcctatatgt gcaaaaaaat gcttgatgat
     2701 ttgtaatgag attgaggagg tttcgagaca ggcaccaaag tttttacaaa tggatatttc
     2761 caccaccgct ctaaccaatt tgttgaagga acacccttgg ctatccttta caagattcga
     2821 actgaagtgg aaacagttgt ctcttatcat ttatgtatta agagattttt tcactaattt
     2881 tacccagaaa aagtcacaac tagaacagga tcaaaatgat catcaaagtt atgaagttaa
     2941 acgatgctcc atcatgttaa gcgatgcagc acaaagaact gttatgtctg taagtagcta
     3001 tatggacaat cataatgtca ccccatattt tgcctggaat tgttcttatt acttgttcaa
     3061 tgcagtccta gtacccataa agactctact ctcaaactca aaatcgaatg ctgagaataa
     3121 cgagaccgca caattattac aacaaattaa cactgttctg atgctattaa aaaaactggc
     3181 cacttttaaa atccagactt gtgaaaaata cattcaagta ctggaagagg tatgtgcgcc
     3241 gtttctgtta tcacagtgtg caatcccatt accgcatatc agttataaca atagtaatgg
     3301 tagcgccatt aaaaatattg tcggttctgc aactatcgcc caatacccta ctcttccgga
     3361 ggaaaatgtc aacaatatca gtgttaaata tgtttctcct ggctcagtag ggccttcacc
     3421 tgtgccattg aaatcaggag caagtttcag tgatctagtc aagctgttat ctaaccgtcc
     3481 accctctcgt aactctccag tgacaatacc aagaagcaca ccttcgcatc gctcagtcac
     3541 gccttttcta gggcaacagc aacagctgca atcattagtg ccactgaccc cgtctgcttt
     3601 gtttggtggc gccaatttta atcaaagtgg gaatattgct gatagctcat tgtccttcac
     3661 tttcactaac agtagcaacg gtccgaacct cataacaact caaacaaatt ctcaagcgct
     3721 ttcacaacca attgcctcct ctaacgttca tgataacttc atgaataatg aaatcacggc
     3781 tagtaaaatt gatgatggta ataattcaaa accactgtca cctggttgga cggaccaaac
     3841 tgcgtataac gcgtttggaa tcactacagg gatgtttaat accactacaa tggatgatgt
     3901 atataactat ctattcgatg atgaagatac cccaccaaac ccaaaaaaag agtaaaatga
     3961 atcgtagata ctgaaaaacc ccgcaagttc acttcaactg tgcatcgtgc accatctcaa
     4021 tttctttcat ttatacatcg ttttgccttc ttttatgtaa ctatactcct ctaagtttca
     4081 atcttggcca tgtaacctct gatctataga attttttaaa tgactagaat taatgcccat
     4141 cttttttttg gacctaaatt cttcatgaaa atatattacg agggcttatt cagaagcttc
     4201 gctcatataa cgaaaaaaaa gggtttggat cgaacgtaat tgagattgat tagttaatac
     4261 tcaaaataaa acagctccta ccaccagtgt aaagtagaac gttaatagag caatgtcttc
     4321 agacaaatct attgagaaaa atacagatac gatcgcctct gaagttcacg aaggtgataa
     4381 tcattcgaat aatttgggtt caatggagga agaggagctc cagaaagcga ccacccaact
     4441 ttggctgata atagcgtata aacaatgcat actttgtacg ttcaaaatac aatgcagtag
     4501 atatatttat gcatattaca tataatacat atcacatagg aagcaacagg cgcgttggac
     4561 ttttaatttt cgaggaccgc gaatccttac atcacaccca atcccccaca agtgatcccc
     4621 cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
     4681 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt
     4741 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
     4801 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg ttctacctga
     4861 atctaagatt gggccctaac tctatcattg atagagtaat tggtatccct atcagtgata
     4921 gagataagcg ggggcggtat ccctatcagt gatagagagg gcttaactct atcattgata
     4981 gagtgccggc aatcctcgag cagattcttt ttcttgaaaa tttttttttt tgattttttt
     5041 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     5101 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     5161 gcatagcaat ctaatctaag ttttaattac aaaactagta aaatggacta caacaagaga
     5221 tcttcggtct caaccgtgcc taatgcagct cccataagag tcggattcgt cggtctcaac
     5281 gcagccaaag gatgggcaat caagacacat taccccgcca tactgcaact atcgtcacaa
     5341 tttcaaatca ctgccttata cagtccaaaa attgagactt ctattgccac catccagcgt
     5401 ctaaaattga gtaatgccac tgcttttccc actttagagt catttgcatc atcttccact
     5461 atagatatga tagtgatagc tatccaagtg gccagtcatt atgacgttgt tatgcctctc
     5521 ttggaattct ccaaaaataa tccgaacctc aagtatcttt tcgtagaatg ggcccttgca
     5581 tgttcactag atcaagccga atccatttat aaggctgctg ctgaacgtgg ggttcaaacc
     5641 atcatctctt tacaaggtcg taaatcacca tatattttga gagcaaaaga attaatatct
     5701 caaggctata tcggcgacat taattctatc gagattgctg gaaatggcgg ttggtacggc
     5761 tacgaaaggc ctgttaaatc accaaaatac atctatgaaa tcgggaacgg tgtagatctg
     5821 gtaaccacaa catttggtca cacaatcgat attttacaat acatgacaag ttcgtacttt
     5881 tccaggataa atgcaatggt ttaaacgcat gcaagcttgg cgtaatcatg gtcatagctg
     5941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8101 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     8161 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     8221 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     8281 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     8341 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     8401 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     8461 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     8521 tcacgacgtt gtaaaacgac ggccagtgaa ttcgtttaaa ccaacctaaa ggtattaact
     8581 tcttcactat aagaaaatca cacgagcgcc cggacgatgt ctctgtttaa atggcgcaag
     8641 ttttccgctt tgtaatatat atttataccc ctttcttctc tcccctgcaa tataatagtt
     8701 taattctaat attaataata tcctatattt tcttcattta ccggcgcact ctcgcccgaa
     8761 cgacctcaaa atgtctgcta cattcataat aaccaaaagc tcataacagg gtaatgaacc
     8821 tgaatatata tacatcatta attccatggg tttagcttgc ctcgtccccg ccgggtcacc
     8881 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     8941 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     9001 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     9061 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     9121 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     9181 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
     9241 ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     9301 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     9361 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     9421 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     9481 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac
     9541 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     9601 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     9661 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     9721 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct
     9781 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     9841 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     9901 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     9961 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
    10021 gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac
    10081 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
    10141 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
    10201 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
    10261 taacgccgcc atccagtgtc gaaaactagg gataacaggg taatgaacct gaatatatat
    10321 acatcattaa ttaatacaat gaaaaacaac accaagctgc caaaagctgt atattaaagt
    10381 taaagctccc tcatttttag taaggacgct ttcttttccc acaaaaggga cgaggaaaaa
    10441 aagaaaaacc tggaaatata gtcacgtgac atgaataaac aagacctagg gcaatatgga
    10501 aaaagaagct aggataaaaa tacatttatg agggttgtaa ggcaaagtgg ctcagcatta
    10561 agtagggcta aaaagagtcg caaatataca tcgtaccgag tgatgaacgt ggttttgaac
    10621 accttgttct cttttgttct cgctccctac attcactata tattagaaga aatttcacct
    10681 caatggacaa ctcgagaaat gaatacagaa atatgttttt tagcgaaatt ttcctttctt
    10741 cttgtcttct tgttttattt aaacttccaa ggctttaact cagtgtcaaa cataacaacc
    10801 tcctcctccc ccacctacga caacaaccgc cactccgcgg atcatgcaga ttttcgtcaa
    10861 gactttgacc ggtaaaacca taacattgga agttgaatct tccgatacca tcgacaacgt
    10921 taagtcgaaa attcaagaca aggaaggtat ccctccagat caacaaagat tgatctttgc
    10981 cggtaagcag ctagaagacg gtagaacgct gtctgattac aacattcaga aggagtccac
    11041 cttacatctt gtgctaaggc taagaggtgg tagacatggt tctggtatca tggttagacc
    11101 attgaactgt atcgttgctg tttctcaaaa catgggtatt ggaaagaatg gtgacttgcc
    11161 ttggccacca ttgagaaatg agtttaagta cttccagaga atgaccacca cttcttcagt
    11221 tgaaggtaaa caaaacttgg tcatcatggg tagaaagacc tggttttcta ttccagaaaa
    11281 gaacaggttg ttgaaggaca gaatcaacat cgttttgtcc agggaattga aagaaccacc
    11341 aagaggtgct catttcttgg caaaatcttt ggatgatgcc ttgagattga tcgaacaacc
    11401 agaattggct tccaaggttg atatggtttg gattgtcggt ggttcatctg tttatcaaga
    11461 agccatgaat caaccaggtc atttgaggtt gttcgttacc agaatcatgc aagagtttga
    11521 gtctgatact ttcttccctg aaattgactt gggcaagtat aagttgttgc cagaatatcc
    11581 aggtgtcttg tctgaagttc aagaagaaaa gggtatcaag tacaagttcg aagtctacga
    11641 aaagaaggat ggtgggggtg gttcttccag attggataag tccaaggtta ttaactctgc
    11701 cttggaattg ttgaacgaag ttggtattga aggtttgacc actagaaagt tggctcaaaa
    11761 attgggtgtt gaacaaccta ccttgtactg gcatgttaag aacaaaagag ctttgttgga
    11821 cgctttggcc attgaaatgt tggatagaca tcatacccat ttctgtccat tggaaggtga
    11881 atcatggcaa gatttcttga gaaacaacgc caagtctttc agatgtgctt tgttgtctca
    11941 tagagatggt gctaaagttc acttgggtac tagaccaact gaaaagcaat acgaaacttt
    12001 ggaaaaccag ttggctttct tgtgtcaaca aggtttctct ttagaaaacg ccttgtatgc
    12061 tttgtctgct gttggtcatt ttaccttggg ttgtgttttg gaagatcaag aacatcaagt
    12121 cgccaaagaa gaaagggaaa ctccaactac tgattctatg ccaccattat tgagacaagc
    12181 cattgagttg tttgatcatc aaggtgctga accagctttt ttgtttggtt tggaattgat
    12241 catctgcggt ttggagaaac agttgaaatg tgaatctggc tcctctacct ctacaactgc
    12301 taaaagaaag aagagaaagt tgactgccag cgtttcgaat acgcagaata agctgaatga
    12361 gcttctcgat gccatcagac aggagtttct ccaagtctca caagaggcaa atacctaccg
    12421 tcttcaaaac caaaaggatt acgatttcaa aatgaaccag cagctggctg agatgcagca
    12481 gataagaaac accgtctacg aactggagct aactcacagg aaaatgaagg acgcgtacga
    12541 agaagagatc aagcacttga aactagggct ggagcaaaga gaccatcaaa ttgcatcttt
    12601 gaccgtccag caacagcagc aacagcaaca gcagcaacag caacagcagc aacaggtcca
    12661 gcagcattta caacagcaac aacagcagct agccgctgca tctgcatctg ttccagttgc
    12721 gcaacaacca ccggctacta cttcggccac cgccactcca gcagcaaaca caactactgg
    12781 ttcgccatcg gccttcccag tacaagctag ccgtcctaat ctggttggct cacagttgcc
    12841 taccaccact ttgcctgtgg tgtcctcaaa cgcccaacaa caactaccac aacagcaact
    12901 gcaacagcag caacttcaac aacagcaacc acctccccag gtttccgtgg caccattgag
    12961 taacacagcc atcaacggat ctcctacttc taaagagacc actactttac cctctgtcaa
    13021 ggcacctgaa tctacgttga aagaaactga accggaaaat aataatacct cgaagataaa
    13081 tgacaccgga tccgccacca cggccaccac taccaccgca actgaaactg aaatcaaacc
    13141 taaggaggaa gacgccaccc cggctagttt gcaccaggat cactacttag tcccttataa
    13201 tcaaagagca aaccactcta aacctatccc acctttcctt ttggatctag attcccagtc
    13261 tgttcccgat gctctgaaga agcaaacaaa tgattattat attttataca acccggcact
    13321 accaagagaa attgacgttg agttacacaa atctttggat catacttcag ttgtttgttg
    13381 cgtgaagttc agtaacgatg gtgaatactt agccacaggc tgcaacaaaa ctactcaagt
    13441 gtatcgcgtt tcagacggtt ctctggtggc ccgtttatct gacgattctg ctgccaataa
    13501 ccatcgaaat tcgatcactg aaaataacac caccacgtcc acggataaca atacaatgac
    13561 aaccactact accaccacaa ttactaccac agcgatgact tcggcagcag aattggcaaa
    13621 agatgtggaa aacctgaaca cttcgtcttc cccatcatcc gacttgtata tccgttcagt
    13681 gtgtttttct ccagatggga aatttttggc aacaggtgct gaagacagac tgattagaat
    13741 ttgggatatt gaaaatagaa agattgttat gattcttcaa ggccacgaac aagatattta
    13801 ttcattggac tactttccct caggtgacaa attagtctcc ggttctggtg accgtaccgt
    13861 tcgtatttgg gacttacgta caggccagtg ttcattgact ttatccattg aagatggtgt
    13921 taccaccgtc gctgtatcac caggtgatgg taaatacatc gctgctggtt ctctagatcg
    13981 tgctgtgaga gtttgggatt ccgagaccgg attcttggtg gaaagactag attcggaaaa
    14041 cgaatccggt acaggccaca aggactctgt ttatagcgtt gtcttcacga gagatggaca
    14101 aagcgttgta tccggctcat tagatagatc tgttaagctc tggaatttgc agaatgcaaa
    14161 caacaagagc gattcgaaaa ctccaaattc cggcacttgt gaagttacgt atatcgggca
    14221 taaagacttt gtattgtccg tggccaccac acaaaatgat gagtacatct tgtccggttc
    14281 caaagatcgt ggtgtcctgt tttgggataa gaaatccggc aatccgttat tgatgttgca
    14341 aggtcatagg aattcagtta tatctgtggc tgtggcaaac gggtctccgc tgggtccaga
    14401 atataacgtt tttgctactg gtagcggtga ttgtaaagca aggatttgga agtataaaaa
    14461 aatagcgcca aattaacctg caggataccc agttgagaag acgagctatt tttctcatgt
    14521 ttcttttgtt tattcacttt tctatacttt ttcttttcta ttgaaatata tattatatct
    14581 tatgtatgtt tttttgtatt ataaacaagc aaatttcgtc ttgccattta ccttct
//
