LOCUS       sequence-194367-        7475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Atoh1 fused to ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  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 7475)
  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
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..50)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        35..364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      215..350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     277..296
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             431..1225
                     /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(485..504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1095..1114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1399..1520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1436..1455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1490..1509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1569..1585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1569..1585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1582..1604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1593..1609
                     /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(1617..1623,1624..1641,1642..1647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1662..1683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1800..1817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1971..2559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2051..2070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2730..3521,3522..3590))
                     /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     3353..3372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3591..3695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3770..3789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3961..4340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4341..4544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4495..4515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4589..4608
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4589..4607
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4684..4693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4693..4740
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     primer_bind     complement(6792..6809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6798..7022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7068..7475,1..21)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7155..7174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7365..7386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
       61 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      181 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
      301 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      361 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
      421 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
      481 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
      541 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
      601 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
      661 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
      721 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
      781 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
      841 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
      901 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
      961 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     1021 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     1081 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     1141 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     1201 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     1261 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     1321 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     1381 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1441 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1501 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     1561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3721 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     3781 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     3841 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     3901 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     3961 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     4021 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4081 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4141 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4201 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4261 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4321 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     4381 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4441 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4501 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
     4561 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
     4621 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
     4681 ttcgccacca tggactacaa agacgatgat gataaagatt acaaagatga cgatgacaag
     4741 gggtcccgcc tgctgcatgc agaagagtgg gctgaggtaa aagagttggg ggaccaccat
     4801 cgccatcccc agccgcacca cgtcccgccg ctgacgccac agccacctgc taccctgcag
     4861 gcgagagacc ttcccgtcta cccggcagaa ctgtccctcc tggatagcac cgacccacgc
     4921 gcctggctga ctcccacttt gcagggcctc tgcacggcac gcgccgccca gtatctgctg
     4981 cattctcccg agctgggtgc ctccgaggcc gcggcgcccc gggacgaggc tgacagccag
     5041 ggtgagctgg taaggagaag cggctgtggc ggcctcagca agagccccgg gcccgtcaaa
     5101 gtacgggaac agctgtgcaa gctgaagggt ggggttgtag tggacgagct tggctgcagc
     5161 cgccagcgag ccccttccag caaacaggtg aatggggtac agaagcaaag gaggctggca
     5221 gcaaacgcaa gggaacggcg caggatgcac gggctgaacc acgccttcga ccagctgcgc
     5281 aacgttatcc cgtccttcaa caacgacaag aagctgtcca aatatgagac cctacagatg
     5341 gcccagatct acatcaacgc tctgtcggag ttgctgcaga ctcccaatgt cggagagcaa
     5401 ccgccgccgc ccacagcttc ctgcaaaaat gaccaccatc accttcgcac cgcctcctcc
     5461 tatgaaggag gtgcgggcgc ctctgcggta gctggggctc agccagcccc gggagggggc
     5521 ccgagaccta ccccgcccgg gccttgccgg actcgcttct caggcccagc ttcctctggg
     5581 ggttactcgg tgcagctgga cgctttgcac ttcccagcct tcgaggacag ggccctaaca
     5641 gcgatgatgg cacagaagga cctgtcgcct tcgctgcccg ggggcatcct gcagcctgta
     5701 caggaggaca acagcaaaac atctcccaga tcccacagaa gtgacggaga gttttccccc
     5761 cactctcatt acagtgactc tgatgaggcc agtctcgagc catctgctgg agacatgagg
     5821 gctgccaacc tttggccaag ccctcttgtg attaagcaca ctaagaagaa tagccctgcc
     5881 ttgtccttga cagctgacca gatggtcagt gccttgttgg atgctgaacc gcccatgatc
     5941 tattctgaat atgatccttc tagacccttc agtgaagcct caatgatggg cttattgacc
     6001 aacctagcag atagggagct ggttcatatg atcaactggg caaagagagt gccaggcttt
     6061 ggggacttga atctccatga tcaggtccac cttctcgagt gtgcctggct ggagattctg
     6121 atgattggtc tcgtctggcg ctccatggaa cacccgggga agctcctgtt tgctcctaac
     6181 ttgctcctgg acaggaatca aggtaaatgt gtggaaggca tggtggagat ctttgacatg
     6241 ttgcttgcta cgtcaagtcg gttccgcatg atgaacctgc agggtgaaga gtttgtgtgc
     6301 ctcaaatcca tcattttgct taattccgga gtgtacacgt ttctgtccag caccttgaag
     6361 tctctggaag agaaggacca catccaccgt gtcctggaca agatcacaga cactttgatc
     6421 cacctgatgg ccaaagctgg cctgactctg cagcagcagc atcgccgcct agctcagctc
     6481 cttctcattc tttcccatat ccggcatatg agtaacaaac gcatggagca tctctacaac
     6541 atgaaatgca agaacgtggt acccctctat gacctgctcc tggagatgtt ggatgcccac
     6601 cgccttcatg ccccagccag tcgcatggga gtgcccccag aggagcccag ccagacccag
     6661 ctggccacca ccagctccac ttcagcacat tccttacaaa cctactacat acccccggaa
     6721 gcagagggct tccccaacac gatctgagcg gccgctcgag tctagagggc ccgtttaaac
     6781 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     7021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     7081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7441 gatttcggcc tattggttaa aaaatgagct gattt
//
