LOCUS       sequence-360204-        7821 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CreOFF tamoxifen-inducible Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7821)
  AUTHORS   Droogers WJ, Willems J, MacGillavry HD, de Jong APH
  TITLE     Duplex Labeling and Manipulation of Neuronal Proteins Using 
            Sequential CRISPR/Cas9 Gene Editing.
  JOURNAL   eNeuro. 2022 Jul 18;9(4):ENEURO.0056-22.2022. doi: 
            10.1523/ENEURO.0056-22.2022.
  PUBMED    35851300
REFERENCE   2  (bases 1 to 7821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/ENEURO.0056-22"; journalName: "eNeuro"; date: "2022-07-18- 
            18"; volume: "9"; issue: "4"; pages: "ENEURO.0056-22.2022"
COMMENT     Sequence Label: pCAG ERT2-Cre-ERT2 lox
FEATURES             Location/Qualifiers
     source          1..7821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1734..1767)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1785..1794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1794..2729
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2739..3761
                     /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="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
                     VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
                     PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
                     IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
                     ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
                     LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
                     D"
     CDS             3774..4709
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             3822..4715
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     protein_bind    complement(4726..4759)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4843..4862)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4908..4963
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4962..4981)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5324..5340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5324..5340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5337..5359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5348..5364
                     /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(5372..5378,5379..5396,5397..5402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5417..5438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5497..5692
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5543..5678
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5698..5832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5735..5754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5789..5808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5900..5917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6071..6659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6151..6170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6830..7621,7622..7690))
                     /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     7453..7472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7691..7795)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrggsggg gsggggsggg gsggggggsg
      541 gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaaggct agcataactt
     1741 cgtatagcat acattatacg aagttatgaa ttcccgggtg agccgccacc atggctggag
     1801 acatgagagc tgccaacctt tggccaagcc cgctcatgat caaacgctct aagaagaaca
     1861 gcctggcctt gtccctgacg gccgaccaga tggtcagtgc cttgttggat gctgagcccc
     1921 ccatactcta ttccgagtat gatcctacca gacccttcag tgaagcttcg atgatgggct
     1981 tactgaccaa cctggcagac agggagctgg ttcacatgat caactgggcg aagagggtgc
     2041 caggctttgt ggatttgacc ctccatgatc aggtccacct tctagaatgt gcctggctag
     2101 agatcctgat gattggtctc gtctggcgct ccatggagca cccagtgaag ctactgtttg
     2161 ctcctaactt gctcttggac aggaaccagg gaaaatgtgt agagggcatg gtggagatct
     2221 tcgacatgct gctggctaca tcatctcggt tccgcatgat gaatctgcag ggagaggagt
     2281 ttgtgtgcct caaatctatt attttgctta attctggagt gtacacattt ctgtccagca
     2341 ccctgaagtc tctggaagag aaggaccata tccaccgagt cctggacaag atcacagaca
     2401 ctttgatcca cctgatggcc aaggcaggcc tgaccctgca gcagcagcac cagcggctgg
     2461 cccagctcct cctcatcctc tcccacatca ggcacatgag taacaaaggc atggagcatc
     2521 tgtacagcat gaagtgcaag aacgtggtgc ccctctatga cctgctgctg gaggcggcgg
     2581 acgcccaccg cctacatgcg cccactagcc gtggaggggc atccgtggag gagacggacc
     2641 aaagccactt ggccactgcg ggctctactt catcgcattc cttgcaaaag tattacatca
     2701 cgggggaggc agagggtttc cctgccacag ctgtcgacaa tttactgacc gtacaccaaa
     2761 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
     2821 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
     2881 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
     2941 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc
     3001 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
     3061 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
     3121 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
     3181 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
     3241 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
     3301 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
     3361 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
     3421 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     3481 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     3541 gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca
     3601 gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca
     3661 tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata
     3721 gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga tctcgagcca tctgctggag
     3781 acatgagagc tgccaacctt tggccaagcc cgctcatgat caaacgctct aagaagaaca
     3841 gcctggcctt gtccctgacg gccgaccaga tggtcagtgc cttgttggat gctgagcccc
     3901 ccatactcta ttccgagtat gatcctacca gacccttcag tgaagcttcg atgatgggct
     3961 tactgaccaa cctggcagac agggagctgg ttcacatgat caactgggcg aagagggtgc
     4021 caggctttgt ggatttgacc ctccatgatc aggtccacct tctagaatgt gcctggctag
     4081 agatcctgat gattggtctc gtctggcgct ccatggagca cccagtgaag ctactgtttg
     4141 ctcctaactt gctcttggac aggaaccagg gaaaatgtgt agagggcatg gtggagatct
     4201 tcgacatgct gctggctaca tcatctcggt tccgcatgat gaatctgcag ggagaggagt
     4261 ttgtgtgcct caaatctatt attttgctta attctggagt gtacacattt ctgtccagca
     4321 ccctgaagtc tctggaagag aaggaccata tccaccgagt cctggacaag atcacagaca
     4381 ctttgatcca cctgatggcc aaggcaggcc tgaccctgca gcagcagcac cagcggctgg
     4441 cccagctcct cctcatcctc tcccacatca ggcacatgag taacaaaggc atggagcatc
     4501 tgtacagcat gaagtgcaag aacgtggtgc ccctctatga cctgctgctg gaggcggcgg
     4561 acgcccaccg cctacatgcg cccactagcc gtggaggggc atccgtggag gagacggacc
     4621 aaagccactt ggccactgcg ggctctactt catcgcattc cttgcaaaag tattacatca
     4681 cgggggaggc agagggtttc cctgccacag cttgatagcg gccgcataac ttcgtatagc
     4741 atacattata cgaagttatg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt
     4801 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     4861 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     4921 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     4981 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5041 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5101 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5161 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5221 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5281 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     5341 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5401 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5461 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     5521 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5581 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5641 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5881 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5941 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6001 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6061 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6121 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6181 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6241 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6301 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6361 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6421 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6481 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6541 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6601 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6661 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6721 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6781 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6961 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7141 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7681 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7741 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7801 ttccccgaaa agtgccacct g
//
