LOCUS       sequence-399602-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid expressing channelrhodopsin-2 fused to a V5 epitope and 
            the ligand-binding domain of estrogen receptor (ChRERa) from a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  AUTHORS   Bonaventura J, Boehm MA, Jedema HP, Solis O, Pignatelli M, Song X, 
            Lu H, Richie CT, Zhang S, Gomez JL, Lam S, Morales M, Gharbawie OA, 
            Pomper MG, Stein EA, Bradberry CW, Michaelides M
  TITLE     Expression of the excitatory opsin ChRERalpha can be traced 
            longitudinally in rat and nonhuman primate brains with PET imaging.
  JOURNAL   Sci Transl Med. 2023 Jul 26;15(706):eadd1014. doi: 
            10.1126/scitranslmed.add1014. Epub 2023 Jul 26.
  PUBMED    37494470
REFERENCE   2  (bases 1 to 7444)
  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: "Sci Transl 
            Med."; date: "2023-07-26"; volume: "15(706):eadd1014. doi"; pages: "
            10.1126/scitranslmed.add1014. Epub 2023 Jul 26"
COMMENT     Sequence Label: pOTTC1755 - pcDNA3.1 CMV-IE hChR2(H134R)-V5-ER...
FEATURES             Location/Qualifiers
     source          1..7444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             920..1846
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1913..1954
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2000..2935
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2048..2941
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     primer_bind     complement(3035..3052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3041..3265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3311..3739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3398..3417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3608..3629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3748..3768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3753..4082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3933..4068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3995..4014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4149..4943
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4203..4222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4813..4832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5117..5238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5154..5173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5208..5227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5287..5303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5287..5303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5300..5322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5311..5327
                     /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(5335..5341,5342..5359,5360..5365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5380..5401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5518..5535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5689..6277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5769..5788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6448..7239,7240..7308))
                     /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     7071..7090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7309..7413)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctaga
      901 cgaagttatc gctagcacca tggactatgg cggcgctttg tctgccgtcg gacgcgaact
      961 tttgttcgtt actaatcctg tggtggtgaa cgggtccgtc ctggtccctg aggatcaatg
     1021 ttactgtgcc ggatggattg aatctcgcgg cacgaacggc gctcagaccg cgtcaaatgt
     1081 cctgcagtgg cttgcagcag gattcagcat tttgctgctg atgttctatg cctaccaaac
     1141 ctggaaatct acatgcggct gggaggagat ctatgtgtgc gccattgaaa tggttaaggt
     1201 gattctcgag ttcttttttg agtttaagaa tccctctatg ctctaccttg ccacaggaca
     1261 ccgggtgcag tggctgcgct atgcagagtg gctgctcact tgtcctgtca tccttatccg
     1321 cctgagcaac ctcaccggcc tgagcaacga ctacagcagg agaaccatgg gactccttgt
     1381 ctcagacatc gggactatcg tgtggggggc taccagcgcc atggcaaccg gctatgttaa
     1441 agtcatcttc ttttgtcttg gattgtgcta tggcgcgaac acattttttc acgccgccaa
     1501 agcatatatc gagggttatc atactgtgcc aaagggtcgg tgccgccagg tcgtgaccgg
     1561 catggcatgg ctgtttttcg tgagctgggg tatgttccca attctcttca ttttggggcc
     1621 cgaaggtttt ggcgtcctga gcgtctatgg ctccaccgta ggtcacacga ttattgatct
     1681 gatgagtaaa aattgttggg ggttgttggg acactacctg cgcgtcctga tccacgagca
     1741 catattgatt cacggagata tccgcaaaac caccaaactg aacatcggcg gaacggagat
     1801 cgaggtcgag actctcgtcg aagacgaagc cgaggccgga gccgtgccag cggccgccac
     1861 cggtggatct atggaagctt actgtcatat taaggggttc gatgccgtga atggaaagcc
     1921 aatacccaat cctctgctgg gcttggactc aactagaacc ggcggcggtt gtccatgggc
     1981 cactggttca ggttcatctg ctggagacat gagagctgcc aacctttggc caagcccgct
     2041 catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg accagatggt
     2101 cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc ctaccagacc
     2161 cttcagtgaa gcctccatga tgggcttact gaccaacctg gcagacaggg agctggttca
     2221 catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc atgatcaggt
     2281 ccaccttctg gaatgtgcct ggctagagat cctgatgatt ggcctcgtct ggcgctccat
     2341 ggagcaccca gggaagctac tgtttgctcc taacttgctc ttggacagga accagggaaa
     2401 atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat ctcggttccg
     2461 catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt tgcttaattc
     2521 tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg accatatcca
     2581 ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg caggcctgac
     2641 cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc acatcaggca
     2701 catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg tggtgcccct
     2761 ctatgacctg ctgctggaga tgctggacgc ccaccgccta catgcgccca ctagccgtgg
     2821 aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct ctacttcatc
     2881 gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg ccacagtcta
     2941 ggcaggtggg cgcgccataa cttcgaattc tgcagatatc cagcacagtg gcggccgctc
     3001 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3061 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3121 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3181 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3241 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3721 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4081 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4141 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4201 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4261 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4321 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4381 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4441 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4501 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4561 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4621 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4681 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4741 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4801 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4861 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4921 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4981 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5041 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5101 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5221 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5881 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5941 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6001 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6061 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7441 cgtc
//
