LOCUS       sequence-477567-        8546 bp    DNA     circular SYN 02-JUL-2025
DEFINITION  Half of photoactivatable GFE3. When co-expressed with PBP-E3-HA 
            (Plasmid #236182), ablates inhibitory synapses after exposure to 400
            nm light..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8546)
  AUTHORS   Bareghamyan A, Deng C, Daoudi S, Yadav SC, Lu X, Zhang W, Campbell 
            RE, Kramer RH, Chenoweth DM, Arnold DB
  TITLE     A toolbox for ablating excitatory and inhibitory synapses.
  JOURNAL   eLife. 2025 Apr 29;13:RP103757. doi: 10.7554/eLife.103757.
  PUBMED    40298577
REFERENCE   2  (bases 1 to 8546)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2025-04-29"; volume: "13:RP103757. doi"; pages: " 
            10.7554/eLife.103757"
COMMENT     Sequence Label: tagRFP-GPHN.FingR-PhoCl
FEATURES             Location/Qualifiers
     source          1..8546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    25..36
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        40..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          697..1713
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1637..1659
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1721..1740
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2137..2844
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     CDS             3658..4383
                     /codon_start=1
                     /product="photocleavable protein that separates into two 
                     fragments upon exposure to violet light (Zhang et al., 
                     2017)"
                     /label=PhoCl
                     /translation="VIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKI
                     HFQGTNFPPNGPVMQKRTVGWEASTEKMYERDGVLKGDVKMKLLLKGGGHYRGDYRTTY
                     KVKQKPVKLPDCHFVDHRIEILSHDKDYNKVKLYEHAVAKTSTDSMDELYKGGSGGMVS
                     KGEETITSVIKPDMKNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFA
                     YDILTTAFHYGNRVFTKYPR"
     intron          4438..4570
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4553..4572
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4579..5167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4632..4652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5050..5061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5194..5211)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5200..5311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5407..5427)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5407..5425)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5446..5462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5568..5587)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5633..5688
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5687..5706)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6049..6065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6073..6089
                     /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(6097..6103,6104..6121,6122..6127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6142..6163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6222..6417
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6268..6403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6423..6557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6460..6479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6514..6533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6625..6642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6796..7384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6876..6895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7555..8346,8347..8415))
                     /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     8178..8197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8416..8520)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagagtcatc ctcatcgtcg acattgatta ttgactagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      481 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      541 sgcgcgccrg gggggggggg gggggggggg gggsggggsg gggsgrggcg grgaggtgcg
      601 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      661 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      721 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      781 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      841 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      901 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      961 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1021 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1081 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1141 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1201 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1261 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggrgggc
     1321 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1381 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1441 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1501 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1561 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1621 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1681 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1741 ctgtctcatc attttggcaa agaattcggt acccttccac agagtatgtt tcagtgccag
     1801 atttgcatgc gcaactttag ccgctcagat acactctcag aacatatacg aacccatacg
     1861 ggagaaaagc cattcgcatg tgatatatgt gggcgcaaat tcgcggctcg aagtacccgg
     1921 acaacccata cgaaaattca cactggttcg caaaagccct ttcagtgtcg aatttgtatg
     1981 cggaatttct cccggtcaga tagtcttagc aaacacatcc gaacacatac gggggaaaag
     2041 ccattcgcct gcgatatttg tggacgaaaa ttcgctcaac gctcaaatct aaaggtacac
     2101 accaaaatcc atctgcgcgg cagccagctg cccggggtgt ctaagggcga agagctgatt
     2161 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
     2221 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
     2281 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
     2341 agcagaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttccct
     2401 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     2461 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     2521 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag
     2581 atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg acatggccct gaagctcgtg
     2641 ggcgggggcc acctgatctg caacttcaag accacataca gatccaagaa acccgctaag
     2701 aacctcaaga tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc
     2761 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
     2821 agcaaactgg ggcacaaact taatggtgga ggttctggag gcggttcagg tggaggttct
     2881 ggaggcggtt ctccatggct cgaagtcaag gaagcatcac caaccagcat ccagatcagc
     2941 tggggcaagt acaaggtcat ggttcgctac taccgcatca cctacggtga aactggtggc
     3001 aatagccctg tccaggaatt caccgtgcct ggcagcaagt ccactgctac catcagcagc
     3061 ctgaaacctg gtgtcgacta taccatcacg gtgtacgccg tcacgatcga ccactggaac
     3121 taccaggacc cgatcccgat ctccatcaac taccgcaccg gttcagctag cggtggaggt
     3181 tctggtggag gttctggagg tggatcagga ggaggttcga gatctaagct ctcaggtggc
     3241 ggagggagcg gcggaggtgg ctctggaggt ggtggctcag ctgaagcatg gtacaatctg
     3301 ggaaacgctt attacaagca gggcgactat caaaaagcca ttgagtatta ccaaaaggct
     3361 cttgagctag accctaataa cgccgaagcg tggtataacc taggcaacgc atattacaag
     3421 cagggagact accaaaaggc aatcgaatat taccagaagg cactcgaact ggacccaaat
     3481 aacgcagagg cttggtataa tctgggaaac gcctactata aacagggtga ctaccaaaag
     3541 gcgatagagg actatcagaa ggccttggag ctcgatccga acaatctgca acgtagtgct
     3601 ggtggcggag ggtctggtgg aggaggctca ggaggaggtg gcgcatcaat cgatatggtg
     3661 atccctgact acttcaagca gagcttcccc gagggctaca gctgggagcg cagcatgacc
     3721 tacgaggacg gcggcatctg catcgccacc aacgacatca caatggaggg ggacagcttc
     3781 atcaacaaga tccacttcca gggcacgaac ttccctccaa acgggcccgt gatgcagaag
     3841 aggaccgtgg gctgggaggc cagcaccgag aagatgtacg agcgcgacgg cgtgctgaag
     3901 ggcgacgtga agatgaagct gctgctgaag ggcggcggcc actatcgcgg cgactaccgc
     3961 accacctaca aggtcaagca gaaaccggta aagctgcccg actgccactt cgtggaccac
     4021 cgcatcgaga tcctgagcca cgacaaggac tacaacaagg tgaagctgta cgagcacgcc
     4081 gtggccaaga cctccaccga cagcatggac gagctgtaca agggtggcag cggtggcatg
     4141 gtgagcaagg gcgaggagac cattacaagc gtgatcaagc ctgacatgaa gaacaagctg
     4201 cgcatggagg gcaacgtgaa cggccacgcc ttcgtgatcg agggcgaggg cagcggcaag
     4261 cccttcgagg gcatccagac gattgatttg gaggtgaagg agggcgcccc gctgcccttc
     4321 gcctacgaca tcctgaccac cgccttccac tacggcaacc gcgtgttcac caagtaccca
     4381 cggacgcgtt aagagagcct gcaattgagt aaagcggccc cggactcgag gccgcaggta
     4441 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     4501 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     4561 ctctccacag gtgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4621 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4681 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4741 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4801 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4861 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4921 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4981 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5041 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5101 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5161 tccccgcctg gaattcgagc tcggtacgat cagcctcgac tgtgccttct agttgccagc
     5221 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5281 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5341 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcccagctt
     5401 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     5461 tgtgtgaaat tgttatccgc taattcactc ctcaggtgca ggctgcctat cagaaggtgg
     5521 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcgatct ttttccctct
     5581 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     5641 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     5701 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     5761 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     5821 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     5881 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     5941 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     6001 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     6061 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6121 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6181 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     6241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     6361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6421 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8521 cacatttccc cgaaaagtgc cacctg
//
