LOCUS       sequence-477270-        8239 bp    DNA     circular SYN 27-JUN-2025
DEFINITION  Fluorescent biosensor for PKA activity proximal to mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  TITLE     Reduced artifacts in nanoscale ratiometric imaging and sensing via 
            PSF matching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3_T20-mSGH-AKAR3EV
FEATURES             Location/Qualifiers
     source          1..8239
                     /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"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1341..1350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1347..2024
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     CDS             2883..3773
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     promoter        complement(3792..3810)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3793..3810)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3830..3847)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3836..4060
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4106..4534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4193..4212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4403..4424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4543..4563)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4548..4877
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4728..4863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4790..4809
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4944..5738
                     /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(4998..5017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5608..5627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5912..6033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5949..5968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6003..6022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6082..6098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6082..6098)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6095..6117)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6106..6122
                     /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(6130..6136,6137..6154,6155..6160))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6175..6196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6313..6330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6484..7072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6564..6583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7243..8034,8035..8103))
                     /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     7866..7885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8104..8208)
                     /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 gctgccacca
      901 tggtgggtcg gaacagcgcc atcgccgccg gtgtatgcgg ggcccttttc attgggtact
      961 gcatctactt cgaccgcaaa agacgaagtg accccaactt caagaacagg cttcgagaac
     1021 gaagaaagaa acagaagctt gccaaggaga gagctgggct ttccaagtta cctgacctta
     1081 aagatgctga agctgttcag aagttcttcc ttgaagaaat acagcttggt gaagagttac
     1141 tagctcaagg tgaatatgag aagggcgtag accatctgac aaatgcaatt gctgtgtgtg
     1201 gacagccaca gcagttactg caggtcttac agcaaactct tccaccacca gtgttccaga
     1261 tgcttctgac taagctccca acaattagtc agagaattgt aagtgctcag agcttggctg
     1321 aagatgatgt ggaagatccc gccaccatgg tgtctacagg cgaggagctg tttaccggcg
     1381 tggtgccctt caagttccag ctgaagggca ccatcaacgg caagagcttc accgtggaag
     1441 gcgagggcga gggcaatagc cacgagggca gccacaaagg caagtacgtg tgcaccagcg
     1501 gcaaactgcc aatgtcttgg gccgccctgg gaactagctt cggctatggc atgaagtact
     1561 acaccaagta ccccagcggc ctgaagaact ggttccacga ggtgatgccc gagggcttca
     1621 cctacgacag acacatccag tacaagggcg acggcagcat ccacgccaag caccagcact
     1681 tcatgaagaa cggcacctac cacaacatcg tggagttcac cggccaggac ttcaaggaga
     1741 acagccccgt gctgaccggc gacatggacg tgagcctgcc caacgaggtg cagcacatcc
     1801 ccatagatga cggcgtggag tgcacagtga ccctgcagta ccctctgctg agcgacgaga
     1861 gcaagtgcgt ggaggcctac cagaacacca tcatcaagcc cctgcacaat cagccagccc
     1921 ccgatgtgcc attccactgg atcagaaagc agtacaccca gagcaaggac gacaccgagg
     1981 agagagacca catcatccag agcgagaccc tggaggccca cctgccatgg cacgagcctt
     2041 ctgcttctgc cgtgatgcat aagttttctc aagaacagat cggcgaaaac attgtgtgca
     2101 gggtcatttg taccacgggt caaattccca tccgagattt gtcagctgat atttcacaag
     2161 tgcttaagga aaaacgatcc ataaagaaag tttggacatt tggtagaaac ccagcctgtg
     2221 actatcattt aggaaacatt tcaagactgt caaataagca tttccaaata ctactaggag
     2281 aagacggtaa ccttttattg aatgacattt ccactaatgg gacctggtta aatgggcaaa
     2341 aagtcgagaa gaacagcaat cagttactgt cccaaggtga tgaaataacc gttggtgtag
     2401 gcgtggaatc agatatttta tctctggtca ttttcataaa cgacaaattt aagcagtgcc
     2461 tggagcagaa caaagttgat cgcggtacca gtgctggtgg tagtgctggt ggtagtgctg
     2521 gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc tggtggtagt gctggtggta
     2581 gtaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     2641 gtggttccgg cagtgctggt ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg
     2701 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt gctggtggta
     2761 gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     2821 gtggtagtgc tggtggttcc ggattgaggc gcgcgacgct ggttgacggc ggcagcaccg
     2881 gtatggcaga aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg
     2941 agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg
     3001 gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc
     3061 gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt
     3121 tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg
     3181 tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc
     3241 cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg
     3301 aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca
     3361 agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc
     3421 cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg
     3481 agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg
     3541 cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct
     3601 ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc
     3661 ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg
     3721 acctgatcgg cagcgagatc gcgcgctggc tgtcgacgct cgagatttcc ggctaatcta
     3781 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3841 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3901 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3961 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4021 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4081 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4141 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4201 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4261 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4321 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4381 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4441 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4501 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4621 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4681 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4921 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4981 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5041 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5101 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5161 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5221 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5281 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5341 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5401 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5461 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5521 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5581 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5641 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5701 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5761 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5821 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5881 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5941 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6001 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6061 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6121 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6181 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6241 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6301 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6361 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6421 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6901 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7021 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7201 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7261 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7321 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7381 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7441 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8041 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8161 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8221 aaaagtgcca cctgacgtc
//
