LOCUS       sequence-182275-        6379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Murine Atoh1 with amino acids 306-347 removed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6379)
  AUTHORS   Cheng YF, Tong M, Edge AS
  TITLE     Destabilization of Atoh1 by E3 Ubiquitin Ligase Huwe1 and Casein 
            Kinase 1 Is Essential for Normal Sensory Hair Cell Development.
  JOURNAL   J Biol Chem. 2016 Sep 30;291(40):21096-21109. Epub 2016 Aug 19.
  PUBMED    27542412
REFERENCE   2  (bases 1 to 6379)
  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: "J Biol 
            Chem."; date: "2016-09-30"; volume: "291(40)"; pages: "21096-21109. 
            Epub 2016 Aug 19"
COMMENT     Sequence Label: FLAG-Atoh1 del4
FEATURES             Location/Qualifiers
     source          1..6379
                     /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             961..1008
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     primer_bind     complement(1970..1987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1976..2200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2246..2674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2333..2352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2543..2564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2683..2703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2688..3017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2868..3003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2930..2949
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3084..3878
                     /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(3138..3157)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3748..3767
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4052..4173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4089..4108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4143..4162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4222..4238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4222..4238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4235..4257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4246..4262
                     /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(4270..4276,4277..4294,4295..4300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4315..4336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4453..4470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4624..5212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4704..4723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5383..6174,6175..6243))
                     /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     6006..6025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6244..6348)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcatg
      961 gactacaaag acgatgatga taaagattac aaagatgacg atgacaaggg gtcccgcctg
     1021 ctgcatgcag aagagtgggc tgaggtaaaa gagttggggg accaccatcg ccatccccag
     1081 ccgcaccacg tcccgccgct gacgccacag ccacctgcta ccctgcaggc gagagacctt
     1141 cccgtctacc cggcagaact gtccctcctg gatagcaccg acccacgcgc ctggctgact
     1201 cccactttgc agggcctctg cacggcacgc gccgcccagt atctgctgca ttctcccgag
     1261 ctgggtgcct ccgaggccgc ggcgccccgg gacgaggctg acagccaggg tgagctggta
     1321 aggagaagcg gctgtggcgg cctcagcaag agccccgggc ccgtcaaagt acgggaacag
     1381 ctgtgcaagc tgaagggtgg ggttgtagtg gacgagcttg gctgcagccg ccagcgagcc
     1441 ccttccagca aacaggtgaa tggggtacag aagcaaagga ggctggcagc aaacgcaagg
     1501 gaacggcgca ggatgcacgg gctgaaccac gccttcgacc agctgcgcaa cgttatcccg
     1561 tccttcaaca acgacaagaa gctgtccaaa tatgagaccc tacagatggc ccagatctac
     1621 atcaacgctc tgtcggagtt gctgcagact cccaatgtcg gagagcaacc gccgccgccc
     1681 acagcttcct gcaaaaatga ccaccatcac cttcgcaccg cctcctccta tgaaggaggt
     1741 gcgggcgcct ctgcggtagc tggggctcag ccagccccgg gagggggccc gagacctacc
     1801 ccgcccgggc cttgccggac tcgcttctca ggcccagctt cctctggggg ttactcggtg
     1861 cagctggacg ctttgcactt cccagccttc gaggacaggg ccctaacagc gatgatggca
     1921 caggatgagg ccagttagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1981 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2041 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2101 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2161 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2221 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2641 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2941 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3001 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3061 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3121 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3181 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3241 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3301 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3361 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3421 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3481 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3541 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3601 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3661 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3721 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3781 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3841 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3901 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3961 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4021 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4201 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6361 aaaagtgcca cctgacgtc
//
