LOCUS       sequence-194369-        8177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Atoh1 fused to ER and DsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8177)
  AUTHORS   Parker MA, Cheng YF, Kinouchi H, Bieber R, Edge AS
  TITLE     An independent construct for conditional expression of atonal 
            homolog-1.
  JOURNAL   Hum Gene Ther Methods. 2014 Feb;25(1):1-13. doi: 
            10.1089/hgtb.2013.014. Epub 2013 Oct 29.
  PUBMED    24066662
REFERENCE   2  (bases 1 to 8177)
  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"; doi: 
            "10.1089/hgtb.2013.014"; journalName: "Hum Gene Ther Methods"; date:
            "2014-02"; volume: "25"; issue: "1"; pages: "1-13"
COMMENT     Sequence Label: CMV-2xFLAG-Atoh1ER-DsRed
FEATURES             Location/Qualifiers
     source          1..8177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(305..733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(415..436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     627..646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    779..1003
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     992..1009
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1054..1731)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1116..1139)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1314..1333)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1531..1551
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1583..1601
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(3763..3810)
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     regulatory      3810..3819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3895..3914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3896..3914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3959..4162)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3988..4008)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4163..4542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4714..4733
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4808..4912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4913..4981,4982..5773)
                     /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     complement(5131..5150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5944..6532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6433..6452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6686..6703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6820..6841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6856..6861,6862..6879,6880..6886)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6894..6910
                     /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)."
     primer_bind     6899..6921
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6918..6934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6918..6934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6983..7104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6994..7013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7048..7067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7278..8072)
                     /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(7389..7408)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7999..8018
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8139..8177,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8153..8177,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
      421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
      481 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
      541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
      601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
      661 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
      721 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
      781 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
      841 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
      901 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
      961 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
     1021 gtttaaacgg gccctctaga ctcgagcggc cgcctacagg aacaggtggt ggcggccctc
     1081 ggcgcgctcg tactgctcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt
     1141 ggagtccacg tagtagtagc cgggcagctg cacgggcttc ttggccatat agatagactt
     1201 gaactccacc aggtagtggc cgccgtcctt cagcttcagg gccttgtgga tctcgccctt
     1261 cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tagtcttctt
     1321 ctgcattacg gggccgtcgg aggggaagtt cacgccgatg aacttcacct tgtagatgaa
     1381 ggagccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     1441 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcttgtagt cggggatgtc
     1501 ggcggggtgc ttcacgtaca ccttggagcc gtactggaac tggggggaca ggatgtccca
     1561 ggcgaagggc agggggccgc ccttggtcac cttcagcttg gcggtctggg tgccctcgta
     1621 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     1681 gcgcaccttg aagcgcatga actccttgat gacgtcctcg gaggaggcca tggttgtggc
     1741 tcctgaacca gatcctgaga tcgtgttggg gaagccctct gcttccgggg gtatgtagta
     1801 ggtttgtaag gaatgtgctg aagtggagct ggtggtggcc agctgggtct ggctgggctc
     1861 ctctgggggc actcccatgc gactggctgg ggcatgaagg cggtgggcat ccaacatctc
     1921 caggagcagg tcatagaggg gtaccacgtt cttgcatttc atgttgtaga gatgctccat
     1981 gcgtttgtta ctcatatgcc ggatatggga aagaatgaga aggagctgag ctaggcggcg
     2041 atgctgctgc tgcagagtca ggccagcttt ggccatcagg tggatcaaag tgtctgtgat
     2101 cttgtccagg acacggtgga tgtggtcctt ctcttccaga gacttcaagg tgctggacag
     2161 aaacgtgtac actccggaat taagcaaaat gatggatttg aggcacacaa actcttcacc
     2221 ctgcaggttc atcatgcgga accgacttga cgtagcaagc aacatgtcaa agatctccac
     2281 catgccttcc acacatttac cttgattcct gtccaggagc aagttaggag caaacaggag
     2341 cttccccggg tgttccatgg agcgccagac gagaccaatc atcagaatct ccagccaggc
     2401 acactcgaga aggtggacct gatcatggag attcaagtcc ccaaagcctg gcactctctt
     2461 tgcccagttg atcatatgaa ccagctccct atctgctagg ttggtcaata agcccatcat
     2521 tgaggcttca ctgaagggtc tagaaggatc atattcagaa tagatcatgg gcggttcagc
     2581 atccaacaag gcactgacca tctggtcagc tgtcaaggac aaggcagggc tattcttctt
     2641 agtgtgctta atcacaagag ggcttggcca aaggttggca gccctcatgt ctccagcaga
     2701 tggctcgaga ctggcctcat cagagtcact gtaatgagag tggggggaaa actctccgtc
     2761 acttctgtgg gatctgggag atgttttgct gttgtcctcc tgtacaggct gcaggatgcc
     2821 cccgggcagc gaaggcgaca ggtccttctg tgccatcatc gctgttaggg ccctgtcctc
     2881 gaaggctggg aagtgcaaag cgtccagctg caccgagtaa cccccagagg aagctgggcc
     2941 tgagaagcga gtccggcaag gcccgggcgg ggtaggtctc gggccccctc ccggggctgg
     3001 ctgagcccca gctaccgcag aggcgcccgc acctccttca taggaggagg cggtgcgaag
     3061 gtgatggtgg tcatttttgc aggaagctgt gggcggcggc ggttgctctc cgacattggg
     3121 agtctgcagc aactccgaca gagcgttgat gtagatctgg gccatctgta gggtctcata
     3181 tttggacagc ttcttgtcgt tgttgaagga cgggataacg ttgcgcagct ggtcgaaggc
     3241 gtggttcagc ccgtgcatcc tgcgccgttc ccttgcgttt gctgccagcc tcctttgctt
     3301 ctgtacccca ttcacctgtt tgctggaagg ggctcgctgg cggctgcagc caagctcgtc
     3361 cactacaacc ccacccttca gcttgcacag ctgttcccgt actttgacgg gcccggggct
     3421 cttgctgagg ccgccacagc cgcttctcct taccagctca ccctggctgt cagcctcgtc
     3481 ccggggcgcc gcggcctcgg aggcacccag ctcgggagaa tgcagcagat actgggcggc
     3541 gcgtgccgtg cagaggccct gcaaagtggg agtcagccag gcgcgtgggt cggtgctatc
     3601 caggagggac agttctgccg ggtagacggg aaggtctctc gcctgcaggg tagcaggtgg
     3661 ctgtggcgtc agcggcggga cgtggtgcgg ctggggatgg cgatggtggt cccccaactc
     3721 ttttacctca gcccactctt ctgcatgcag caggcgggac cccttgtcat cgtcatcttt
     3781 gtaatcttta tcatcatcgt ctttgtagtc catggtggcg aattccacca cactggacta
     3841 gtggatccga gctcggtacc aagcttaagt ttaaacgcta gccagcttgg gtctccctat
     3901 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     3961 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     4021 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     4081 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     4141 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4201 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4261 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4321 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4381 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4441 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4501 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc
     4561 gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg
     4621 tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca
     4681 cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag
     4741 tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg
     4801 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4861 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4921 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4981 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5041 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5101 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5161 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5221 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5281 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5341 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5401 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5461 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5521 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5581 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5641 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5701 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5761 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6721 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6781 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     6841 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     6901 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     6961 ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     7021 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     7081 attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga gatccccgcg
     7141 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     7201 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     7261 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     7321 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     7381 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     7441 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     7501 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     7561 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     7621 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     7681 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     7741 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     7801 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     7861 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     7921 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     7981 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     8041 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     8101 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     8161 aaaaagcctc ctcacta
//
