LOCUS       sequence-380404-       12395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral production of HIF-1alpha reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12395)
  AUTHORS   Jeknic S, Kudo T, Song JJ, Covert MW
  TITLE     An optimized reporter of the transcription factor hypoxia-inducible 
            factor 1alpha reveals complex HIF-1alpha activation dynamics in 
            single cells.
  JOURNAL   J Biol Chem. 2023 Mar 10:104599. doi: 10.1016/j.jbc.2023.104599.
  PUBMED    36907438
REFERENCE   2  (bases 1 to 12395)
  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: "J Biol 
            Chem. 2023 Mar 10:104599. doi: 10.1016/j.jbc.2023.104599."
COMMENT     Sequence Label: pLenti-HIFR
FEATURES             Location/Qualifiers
     source          1..12395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(210..1001,1002..1070))
                     /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     833..852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1071..1175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1250..1269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1441..1820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1822..2020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1975..1995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2038..2218
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2265..2390
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2887..3120
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3305..3349
                     /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    3647..3764
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    3835..3859
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3865..4400
                     /label=mCMV promoter
                     /note="mouse cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             5942..6649
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6197..6206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7934..7990
                     /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             7991..8368
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             join(8423..8425,8426..8428,8429..9139,9140..9145,
                     9146..9268)
                     /codon_start=1
                     /product="ECFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(8468..8489)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8729..8748)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9076..9097
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(9292..9316)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    9344..9932
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9397..9417)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9815..9826)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9935..9951)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9936..9952)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        10083..10570
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10118..10141)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     10503..10522
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             10614..11213
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(10614..10633)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     11110..11130
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             11604..11784
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(join(11846..12395,1..39))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11926..11945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
       61 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
      661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
      721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
      781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
      841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
      901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
      961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1201 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     1261 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     1321 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     1381 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     1441 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     1501 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     1561 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     1621 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1681 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1741 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1801 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     1861 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     1921 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     1981 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
     2041 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     2101 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     2161 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     2221 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     2281 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     2341 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     2401 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     2461 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     2521 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     2581 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     2641 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     2701 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     2761 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     2821 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     2881 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     2941 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     3001 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     3061 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     3121 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     3181 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     3241 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     3301 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     3361 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     3421 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     3481 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     3541 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     3601 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     3661 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     3721 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     3781 acagcagaga tccagtttgg ttagtaccgg gccggtggaa ttctgcagat atcaacaagt
     3841 ttgtacaaaa aagcaggctt cgagtggcac gtatactgag tcattaggga ctttccaatg
     3901 ggttttgccc agtacataag gtcaataggg gtgaatcaac aggaaagtcc cattggagcc
     3961 aagtacactg agtcaatagg gactttccat tgggttttgc ccagtacaaa aggtcaatag
     4021 ggggtgagtc aatgggtttt tcccattatt ggcacgtaca taaggtcaat aggggtgagt
     4081 cattgggttt ttccagccaa tttaattaaa acgccatgta ctttcccacc attgacgtca
     4141 atgggctatt gaaactaatg caacgtgacc tttaaacggt actttcccat agctgattaa
     4201 tgggaaagta ccgttctcga gccaatacac gtcaatggga agtgaaaggg cagccaaaac
     4261 gtaacaccgc cccggttttc ccctggaaat tccatattgg cacgcattct attggctgag
     4321 ctgcgttcta cgtgggtata agaggcgcga ccagcgtcgg taccgtcgca gtcttcggtc
     4381 tgaccaccgt agaacgcaga gtgatccacc ggggacgcga ggactgtcct cgccgccgtc
     4441 gcgggcagtg tctagccagg ccttgacaag ctagccggag gagcgcctag gaacccgagc
     4501 cggagctcag cgagcgcagc ctgcagctcc cgcctcgccg tcccgggggg cgtcccgcct
     4561 cccaccccgc ctctggactt gtctctttct ccgcgcgcgc ggacacagcc ggcgcttagg
     4621 cccgagcgag cccgggggcc gccggccggg aagacaacgc gggcaccgat tcgccatgga
     4681 gggcgccggc ggcgagaacg agaagaaaaa gatgagttct gaacgtcgaa aagaaaagtc
     4741 tagagatgca gcaagatctc ggcgaagcaa agagtctgaa gttttttatg agcttgctca
     4801 tcagttgcca cttccccaca atgtgagctc acatcttgat aaagcttctg ttatgaggct
     4861 caccatcagt tatttacgtg tgagaaaact tctggatgcc ggtggtctag acagtgaaga
     4921 tgagatgaag gcacagatgg actgttttta tctgaaagcc ctagatggct ttgtgatggt
     4981 gctaacagat gacggcgaca tggtttacat ttctgataac gtgaacaaat acatggggtt
     5041 aactcagttt gaactaactg gacacagtgt gtttgatttt actcatccat gtgaccatga
     5101 ggaaatgaga gaaatgctta cacacagaaa tggcccagtg agaaaaggga aagaactaaa
     5161 cacacagcgg agcttttttc tcagaatgaa gtgcacccta acaagccggg ggaggacgat
     5221 gaacatcaag tcagcaacgt ggaaggtgct tcactgcacg ggccatattc atgtctatga
     5281 taccaacagt aaccaacctc agtgtgggta caagaaacca cccatgacgt gcttggtgct
     5341 gatttgtgaa cccattcctc atccgtcaaa tattgaaatt cctttagaca gcaagacatt
     5401 tctcagtcga cacagcctcg atatgaaatt ttcttactgt gatgaaagaa ttactgagtt
     5461 gatgggttat gagccggaag aacttttggg ccgctcaatt tatgaatatt atcatgcttt
     5521 ggattctgat catctgacca aaactcacca tgatatgttt actaaaggac aagtcaccac
     5581 aggacagtac aggatgcttg ccaaaagagg tggatatgtc tgggttgaaa ctgaagcaac
     5641 tgtcatatat aatacgaaga actcccagcc acagtgcatt gaatgtgtga atactgttgt
     5701 aagtggtatt attcagcacg acttgatttt ctcccttcaa caaacagaat ctgtgctcaa
     5761 accagttgaa tcttcagata agaagatgac tcagctgttc accaaagttg aatcagaggc
     5821 tacaagctgc ctttttgata agcttaagaa ggagcctgat gctctcactc tgctggctcc
     5881 agctgccggc gacaccatca tctctctgga ttttggcagc gatgacacag aaggttccgg
     5941 catggtgagc aagggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca
     6001 catctttggc tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc
     6061 aaatgatggt tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc
     6121 ctggattctg gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg
     6181 gatgtcgcct ttccaggccg ccatggtaga tggctccggc taccaagtcc atcgcacaat
     6241 gcagtttgaa gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca
     6301 catcaaagga gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac
     6361 caactcgctg accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac
     6421 catcatcagt acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac
     6481 tgcgcggacc acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc
     6541 gatgtacgtg ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga
     6601 gtggcaaaag gcctttaccg atgtgatggg catggacgag ctgtacaagg gctcaggaac
     6661 tgaagatcaa caacttgaag atgttccatt atataatgat gtaatgtttc cctcttctaa
     6721 tgaaaaatta aatataaacc tggcaatgtc tcctttacct tcatcggaaa ctccaaagcc
     6781 acttcgaagt agtgctgatc ctgcacttaa tcaagaggtt gcattaaaat tagaatcaag
     6841 tccagagtca ctgggacttt cttttaccat gccccagatt caagatcagc cagcaagtcc
     6901 ttctgatgga agcactagac aaagttcacc tgagagactt cttcaggaaa acgtaaacac
     6961 tcctaacttt tcccagccta acagtcccag tgaatattgc tttgatgtgg atagcgatat
     7021 ggtcaatgta ttcaagttgg aactggtgga aaaactgttt gctgaagaca cagaggcaaa
     7081 gaatccattt tcaactcagg acactgattt agacttggag atgctggctc cctatatccc
     7141 aatggatgat gatttccagt tacgttcctt tgatcagttg tcaccattag agagcaattc
     7201 tccaagccct ccaagtatga gcacagttac tgggttccag cagacccagt tacagaaacc
     7261 taccatcact gccactgcca ccacaactgc caccactgat gaatcaaaaa cagagacgaa
     7321 ggacaataaa gaagatatta aaatactgat tgcatctcca tcttctaccc aagtacctca
     7381 agaaacgacc actgctaagg catcagcata cagtggcact cacagtcgga cagcctcacc
     7441 agacagagca ggaaagagag tcatagaaca gacagacaaa gctcatccaa ggagccttaa
     7501 cctgtctgcc actttgaatc aaagaaatac tgttcctgag gaagaattaa acccaaagac
     7561 aatagcttcg cagaatgctc agaggaagcg aaaaatggaa catgatggct ccctttttca
     7621 agcagcagga attggaacat tattgcagca accgggtgac tgtgcaccta ctatgtcact
     7681 ttcctggaaa cgagtgaaag gattcatatc tagtgaacag aatggaacgg agcaaaagac
     7741 tattatttta ataccctccg atttagcatg cagactgctg gggcagtcaa tggatgagag
     7801 tggattacca caggcaacca gttacgattg tgaagttaat gctcccatac aaggcagcag
     7861 aaacctactg cagggtgaag aattactcag agctttggat caagttaacg gtggcggtgg
     7921 cagcggaagc ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa
     7981 ccctggacct atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa
     8041 ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag
     8101 ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc
     8161 caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga
     8221 ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac
     8281 ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa
     8341 ggccatcacc aagtacacca gcgctaagaa catggggcat ggcaccggca gcaccggcag
     8401 cggcagctca tccggaacta gtatggtgag caagggcgag gagctgttca ccggggtggt
     8461 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     8521 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     8581 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc
     8641 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     8701 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     8761 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     8821 ggacggcaac atcctggggc acaagctgga gtacaacgcc atcagcgata acgtctatat
     8881 caccgccgac aagcagaaga acggcatcaa ggccaacttc aaaattagac acaacatcga
     8941 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     9001 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     9061 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     9121 catggacgag ctgtacaaga agcttagcca tggcttcccg ccggaggtgg aggagcagga
     9181 tgatggcacg ctgcccatgt cttgtgccca ggagagcggg atggaccgtc accctgcagc
     9241 ctgtgcttct gctaggatca atgtgtaagc ggccgcactc gagatatcta gacccagctt
     9301 tcttgtacaa agtggttgat atccagcaca gtggcggccc gataatcaac ctctggatta
     9361 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     9421 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     9481 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     9541 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     9601 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     9661 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     9721 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     9781 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     9841 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     9901 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc
     9961 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
    10021 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacactcagg tacaattcta
    10081 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
    10141 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
    10201 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
    10261 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
    10321 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
    10381 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
    10441 gggtgggtcc gggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
    10501 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
    10561 gggcctttcg acctgcatcc atctagatct cgatcgagca gctgaagctt accatgaccg
    10621 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
    10681 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
    10741 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
    10801 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
    10861 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
    10921 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
    10981 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
    11041 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
    11101 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
    11161 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtacctt
    11221 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
    11281 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
    11341 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
    11401 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
    11461 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
    11521 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
    11581 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
    11641 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
    11701 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    11761 agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc
    11821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    11881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    11941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    12001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    12061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    12121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    12181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    12241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    12301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    12361 aacaaaccac cgctggtagc ggtggttttt ttgtt
//
