LOCUS       sequence-194359-        6505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Atoh1 with FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6505)
  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 6505)
  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-Atoh1
FEATURES             Location/Qualifiers
     source          1..6505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(225..653)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(335..356)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     547..566
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    699..923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     912..929
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2017..2064)
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     primer_bind     complement(2143..2162)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2144..2162)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2207..2410)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2236..2256)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2411..2790
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2962..2981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3056..3160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3161..3229,3230..4021)
                     /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(3379..3398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4192..4780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4681..4700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4934..4951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5068..5089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5104..5109,5110..5127,5128..5134)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5142..5158
                     /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     5147..5169
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5166..5182
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5166..5182
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5231..5352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5242..5261)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5296..5315
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5526..6320)
                     /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(5637..5656)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6247..6266
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6387..6505,1..211))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6401..6505,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6455..6474)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt
      241 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      301 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      361 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      421 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      481 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      541 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      601 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtggggata
      661 ccccctagag ccccagctgg ttctttccgc ctcagaagcc atagagccca ccgcatcccc
      721 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
      781 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
      841 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      901 tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gtttaaacgg gccctctaga
      961 ctaactggcc tcatcagagt cactgtaatg agagtggggg gaaaactctc cgtcacttct
     1021 gtgggatctg ggagatgttt tgctgttgtc ctcctgtaca ggctgcagga tgcccccggg
     1081 cagcgaaggc gacaggtcct tctgtgccat catcgctgtt agggccctgt cctcgaaggc
     1141 tgggaagtgc aaagcgtcca gctgcaccga gtaaccccca gaggaagctg ggcctgagaa
     1201 gcgagtccgg caaggcccgg gcggggtagg tctcgggccc cctcccgggg ctggctgagc
     1261 cccagctacc gcagaggcgc ccgcacctcc ttcataggag gaggcggtgc gaaggtgatg
     1321 gtggtcattt ttgcaggaag ctgtgggcgg cggcggttgc tctccgacat tgggagtctg
     1381 cagcaactcc gacagagcgt tgatgtagat ctgggccatc tgtagggtct catatttgga
     1441 cagcttcttg tcgttgttga aggacgggat aacgttgcgc agctggtcga aggcgtggtt
     1501 cagcccgtgc atcctgcgcc gttcccttgc gtttgctgcc agcctccttt gcttctgtac
     1561 cccattcacc tgtttgctgg aaggggctcg ctggcggctg cagccaagct cgtccactac
     1621 aaccccaccc ttcagcttgc acagctgttc ccgtactttg acgggcccgg ggctcttgct
     1681 gaggccgcca cagccgcttc tccttaccag ctcaccctgg ctgtcagcct cgtcccgggg
     1741 cgccgcggcc tcggaggcac ccagctcggg agaatgcagc agatactggg cggcgcgtgc
     1801 cgtgcagagg ccctgcaaag tgggagtcag ccaggcgcgt gggtcggtgc tatccaggag
     1861 ggacagttct gccgggtaga cgggaaggtc tctcgcctgc agggtagcag gtggctgtgg
     1921 cgtcagcggc gggacgtggt gcggctgggg atggcgatgg tggtccccca actcttttac
     1981 ctcagcccac tcttctgcat gcagcaggcg ggaccccttg tcatcgtcat ctttgtaatc
     2041 tttatcatca tcgtctttgt agtccatgaa ttccaccaca ctggactagt ggatccgagc
     2101 tcggtaccaa gcttaagttt aaacgctagc cagcttgggt ctccctatag tgagtcgtat
     2161 taatttcgat aagccagtaa gcagtgggtt ctctagttag ccagagagct ctgcttatat
     2221 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     2281 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     2341 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     2401 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     2461 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     2521 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     2581 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     2641 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     2701 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     2761 actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa
     2821 gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc
     2881 ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga
     2941 gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg
     3001 gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg
     3061 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4321 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4921 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4981 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5041 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5101 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5161 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat
     5221 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5281 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5341 aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     5401 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     5461 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     5521 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     5581 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     5641 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     5701 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     5761 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     5821 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     5881 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     5941 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     6001 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     6061 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     6121 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     6181 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     6241 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     6301 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatccgaaa
     6361 atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct
     6421 cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa
     6481 attagtcagc catggggcgg agaat
//
