LOCUS       sequence-478721-        5781 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  inducible mouse Adar Deaminase domain (HyperTRIBE) for expression in
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  TITLE     Stemness factor Mex3a times translation and protein trafficking to 
            ensure robust differentiation of olfactory sensory neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: tetO-HyperTRIBE-adar_deaminase-V5-t2a-mCherry
FEATURES             Location/Qualifiers
     source          1..5781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(20..38,39..61,62..80,81..103,104..122,123..145,
                     146..164,165..187,188..206,207..229,230..248,249..271,
                     272..290)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        323..361
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     358..382
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1718..1759
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1781..1834
                     /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             1841..2548
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1983..2001)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2251..2270
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2570..3142
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3274..3293)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3339..3394
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3393..3412)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3792..3809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3963..4551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4043..4062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4722..5513,5514..5582))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5345..5364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5583..5687)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5755..5773
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcactcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
       61 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
      121 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
      181 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      241 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      301 tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc
      361 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      421 aagacaccgg gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc
      481 ctctagtcag ctgacgcgtg ctagcgcggc catgtccggc cgcatcgata agcttggagg
      541 tggaggtagt accttccacg accagatagc tatgctgagc cacaggtgct tcaatgctct
      601 gaccaacagt ttccagccct ccctgctcgg ccgcaagatc ctggctgcca ttattatgaa
      661 aagagatcca gaggacatgg gtgttgtcgt gagtttgggg acaggcaatc gctgtgtgaa
      721 aggggactct ctcagcctga agggagagac ggtcaatgac tgccatgccg aaatcatctc
      781 ccggaggggc ttcatcaggt ttctctacag tgaactgatg aagtacaacc accacactgc
      841 caagaacagc atatttgagc ttgccagggg aggagagaag ctgcagataa agaagacggt
      901 ttcttttcat ctctacatca gcacggcgcc atgtggagat ggggccctct ttgacaaatc
      961 ctgcagtgac cgtgccgtgg aaagcacaga gtcccgccat taccctgtct ttgaaaatcc
     1021 caagcaaggc aagcttcgca ccaaggtgga gaatgggcaa ggcacaattc ctgtggagtc
     1081 cagtgatatt gtacccacgt gggatggcat ccgccttggg gaaagactcc gtaccatgtc
     1141 ctgtagtgac aaaatcctac gctggaatgt gctgggcctg caaggggcgt tgttgacgca
     1201 cttcctacag cctgtgtacc tgaaatctgt aaccttaggt taccttttca gccaagggca
     1261 tctgacccgt gctatttgct gccgcgtgac cagagatggg aaagcatttg aggatggact
     1321 acgctatccc tttattgtca accaccccaa ggtcggccga gtcagtgttt atgattccaa
     1381 aagacagtcc ggaaagacca aggaaacaag cgtcaactgg tgcatggctg atggctatga
     1441 cctagagatc ctggatggca ccagaggcac tgtggatgga ccagggaaag agttgtctcg
     1501 ggtgtccaag aagaatattt tccttcagtt taagaagctc tgctccttcc gagcccgcag
     1561 agatttactg cagctctctt atggtgaagc caagaaagct gcccgtgact atgacttagc
     1621 caagaactac ttcaagaaaa gcctgcgaga catgggctat gggaattgga tcagcaaacc
     1681 ccaggaggaa aagaacttta cggatcccca gccatggggt aagcctatcc ctaaccctct
     1741 cctcggtctc gattctacga agcttgtcga cggcagtgga gagggcagag gaagtctgct
     1801 aacatgcggt gacgtcgagg agaatcctgg cccatctaga gtgagcaagg gcgaggagga
     1861 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     1921 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1981 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2041 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2101 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2161 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2221 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2281 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2341 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2401 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2461 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2521 cggcggcatg gacgagctgt acaagtaatc tagagctgag aacttcaggg tgagtttggg
     2581 gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa
     2641 agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg
     2701 ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt
     2761 ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt
     2821 cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa
     2881 tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg
     2941 ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag
     3001 aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat gatatacact
     3061 gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
     3121 tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     3181 ttttggcaaa gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3241 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3301 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3361 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3421 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3481 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3541 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3601 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3661 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     3721 cctcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4861 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     5341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5701 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5761 cgtatcacga ggccctttcg t
//
