LOCUS       sequence-343756-        7515 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Co-expression of HaloTag11 (actin) and 7 (mitochondrial surface) in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7515)
  AUTHORS   Frei MS, Tarnawski M, Roberti MJ, Koch B, Hiblot J, Johnsson K
  TITLE     Engineered HaloTag variants for fluorescence lifetime multiplexing.
  JOURNAL   Nat Methods. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi: 
            10.1038/s41592-021-01341-x.
  PUBMED    34916672
REFERENCE   2  (bases 1 to 7515)
  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"; journalName: "Nat 
            Methods. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi: 
            10.1038/s41592-021-01341-x."
COMMENT     Sequence Label: pcDNA5/FRT/TO_Lifeact-HaloTag11-T2A-Tomm20-Hal...
FEATURES             Location/Qualifiers
     source          1..7515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    697..715
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    718..736
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     746..770
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             920..970
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             998..1882
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPWGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1898..1951
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2417..3301
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3344..3361)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3350..3574
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3707..3726)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3858..3905
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3913..4935
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3913..4935
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5065..5186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5102..5121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5156..5175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5235..5251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5235..5251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5248..5270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5259..5275
                     /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(5283..5289,5290..5307,5308..5313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5328..5349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5466..5483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5637..6225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5717..5736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6396..7187,7188..7256))
                     /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     7019..7038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7257..7361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7436..7455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
       61 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc
      721 ctatcagtga tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac
      781 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      841 tagcgtttaa acttaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt
      901 ctgcagatat ccagcaacca tgggcgtggc cgacctgatc aagaagttcg agagcatcag
      961 caaggaggag ggggatccac cggtcgccac cggatccgaa atcggtactg gctttccatt
     1021 cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg
     1081 cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa
     1141 catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg
     1201 caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc
     1261 cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc
     1321 tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga
     1381 gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca
     1441 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
     1501 gggtacgctg ccgtggggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
     1561 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
     1621 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
     1681 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
     1741 cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg
     1801 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     1861 gtctactctg gagatttccg gtggccgcgg cagtggagag ggcagaggaa gtctgctaac
     1921 atgcggtgac gtcgaggaga atcctggccc aatggtgggt cggaacagcg ccatcgccgc
     1981 cggtgtatgc ggggcccttt tcattgggta ctgcatctac ttcgaccgca aaagacgaag
     2041 tgaccccaac ttcaagaaca ggcttcgaga acgaagaaag aaacagaagc ttgccaagga
     2101 gagagctggg ctttccaagt tacctgacct taaagatgct gaagctgttc agaagttctt
     2161 ccttgaagaa atacagcttg gtgaagagtt actagctcaa ggtgaatatg agaagggcgt
     2221 agaccatctg acaaatgcaa ttgctgtgtg tggacagcca cagcagttac tgcaggtctt
     2281 acagcaaact cttccaccac cagtgttcca gatgcttctg actaagctcc caacaattag
     2341 tcagagaatt gtaagtgctc agagcttggc tgaagatgat gtggaaggcg gtagcgggga
     2401 tccaccggtc gccaccgaga ttggcacagg gtttcccttt gaccctcatt acgttgaggt
     2461 gctgggtgag aggatgcact atgtcgatgt gggaccccgt gatggcacac ctgtgctgtt
     2521 tctccatggc aatcctacca gctcctatgt gtggaggaac atcataccgc atgtggctcc
     2581 aacgcaccga tgtatcgcac cagatctgat cgggatggga aagagcgaca aacctgatct
     2641 gggatacttc ttcgatgacc acgtgcgctt catggacgcc tttatcgagg ctttggggct
     2701 tgaggaggta gtgctggtca tccacgattg gggatctgcc ctggggtttc attgggcgaa
     2761 acgcaatcct gagagggtta agggcatagc cttcatggag ttcatacggc ccattccaac
     2821 atgggacgaa tggccggaat ttgctcggga aacctttcaa gcgttccgca ctaccgatgt
     2881 tgggagaaag ctcatcatcg accaaaacgt gttcatcgaa ggaactctcc cgatgggtgt
     2941 ggtacgaccc ttgactgaag tcgagatgga ccactataga gagccgtttc tcaacccagt
     3001 ggacagagaa cctctgtgga ggttccccaa cgaacttccc attgcaggcg aaccagccaa
     3061 tatcgtcgcc ctcgttgagg agtacatgga ctggttgcat cagtcaccag tacccaagct
     3121 gttgttctgg ggtacaccag gagtgctgat tcctcccgca gaagcagctc ggctggctaa
     3181 gtctcttccc aactgcaaag ccgtcgacat tggcccaggt ctgaatctgc ttcaggagga
     3241 taatcctgat ctgattggca gtgagattgc cagatggctg tcaaccctgg aaatctccgg
     3301 taaccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3361 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3421 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3481 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3541 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3601 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3661 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3721 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3781 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3841 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     3901 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     3961 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4021 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4081 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4141 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4201 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4261 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4321 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4381 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4441 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4501 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     4561 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     4621 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     4681 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     4741 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     4801 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     4861 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     4921 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4981 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5041 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5101 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5161 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6061 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7381 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7441 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7501 ctgagtagtg cgcga
//
