LOCUS       sequence-365047-        4689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible expression of constitutively active Retinoic 
            Acid Receptor alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4689)
  AUTHORS   Amamoto R, Wallick GK, Cepko CL
  TITLE     Retinoic acid signaling mediates peripheral cone photoreceptor 
            survival in a mouse model of retina degeneration.
  JOURNAL   Elife. 2022 Mar 22;11. pii: 76389. doi: 10.7554/eLife.76389.
  PUBMED    35315776
REFERENCE   2  (bases 1 to 4689)
  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: "Elife."; 
            date: "2022-03-22"; volume: "11. pii: 76389. doi"; pages: " 
            10.7554/eLife.76389"
COMMENT     Sequence Label: TRE-RARa-VP16
FEATURES             Location/Qualifiers
     source          1..4689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        125..439
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    169..187
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    204..222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    240..258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    276..294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    311..329
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    347..365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     412..436
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      445..454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1987..2220
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    2259..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2324..2343
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2628..2645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2799..3387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2879..2898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3587..4378,4379..4447))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4552..4600
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4614..4689,1..16)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4654..4673
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gccagaacat ttctctggcc taactggccg gtacctgagc tcgctagcct cgaggatatc
       61 aagatctggc ctcggcggcc aagcttgcat gcctgcaggt cacgaggccc tttcgtcttc
      121 actcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      181 atagagaacg atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
      241 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat
      301 gtcgagttta tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      361 agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      421 agtgaaccgt cagatcgcct ggaggccacc atggccagca atagcagttc ctgcccaaca
      481 cctgggggcg ggcacctcaa tgggtaccca gtacccccct acgccttctt ctttccccct
      541 atgctgggtg gactctcccc acccggcgct ctcaccagcc tccagcacca gcttccagtc
      601 agtggttaca gcacaccgtc cccagccacc atcgagaccc agagcagcag ttccgaagag
      661 atagtaccca gccctccctc accaccgccc ctgccccgca tctacaagcc ttgctttgtt
      721 tgtcaagaca aatcatccgg ctaccactat ggggtcagcg cctgtgaggg ctgtaagggc
      781 ttcttccgac gaagcatcca gaagaacatg gtgtatacgt gtcaccggga caagaactgc
      841 atcatcaaca aggtgacccg gaaccgctgc cagtactgcc ggctgcagaa atgtttcgac
      901 gtgggcatgt ccaaggagtc ggtgcgaaac gatcgaaaca aaaagaagaa agaggcaccc
      961 aagcccgagt gctcagagag ctacacgctg acgcctgagg tgggcgagct cattgagaag
     1021 gttcgcaaag cgcaccagga gaccttcccg gccctctgcc agctgggcaa gtacactacg
     1081 aacaacagct cagaacaacg agtctccctg gacattgacc tctgggacaa gttcagtgaa
     1141 ctctccacca agtgcatcat taagactgtg gagttcgcca agcagcttcc cggcttcacc
     1201 accctcacca tcgccgacca gatcaccctc ctcaaggctg cctgcctgga tatcctgatt
     1261 ctgcgaatct gcacgcggta cacgcctgag caagacacaa tgaccttctc agatggactg
     1321 accctgaacc ggactcagat gcacaacgct ggctttggcc ccctcaccga cttggtcttt
     1381 gccttcgcca accagctgct gcccctggag atggacgatg ctgagactgg actgctcagt
     1441 gccatctgcc tcatctgtgg agaccgacag gacctggagc agccagacaa ggtggacatg
     1501 ctgcaagagc cgctgctgga agcactgaaa gtctacgtcc ggaaacggag gcccagccga
     1561 ccccacatgt tccccaagat gctgatgaag atcacagacc ttcggagcat cagcgccaag
     1621 ggagctgaac gggtgatcac attgaagatg gagatcccag gctccatgcc accgctgatc
     1681 caggaaatgc tggagaactc tgagggcttg gacactctaa gcggacagtc ggggggcgga
     1741 acacgagatg ggggtggcct ggccccccct ccgggtagct gtagccccag cctcagtccc
     1801 agctcccaca gaagcagccc agccacccaa tccccaggac cggtccgcgc gcgtacgaaa
     1861 aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga cgcccccgaa
     1921 gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac gcgcagactg
     1981 tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
     2041 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
     2101 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
     2161 gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg
     2221 taggagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2281 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2341 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2401 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2461 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2521 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2581 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3541 taaacttggt ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga
     3601 tcagtgaggc accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc
     3661 ccgtcgtgta gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga
     3721 tgccgcgaga gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga
     3781 gggccgagcg aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct
     3841 gtcgtgatgc tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg
     3901 ctactggcat cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc
     3961 agcggtcaag ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag
     4021 ggcctccgat cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag
     4081 cactacacaa ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt
     4141 actcaaccaa gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt
     4201 ctatacggga caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc
     4261 gttcttcggg gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc
     4321 ccactcttgc acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg
     4381 caaaaacagg caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga
     4441 tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct
     4501 ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata
     4561 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     4621 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     4681 agtgcaggt
//
