LOCUS       sequence-274396-        5141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Six1 with a myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5141)
  AUTHORS   Taylor-Weiner H, Grigsby CL, Ferreira DMS, Dias JM, Stevens MM, Ruas
            JL, Teixeira AI
  TITLE     Modeling the transport of nuclear proteins along single skeletal 
            muscle cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Feb 11;117(6):2978-2986. doi: 
            10.1073/pnas.1919600117. Epub 2020 Jan 27.
  PUBMED    31988126
REFERENCE   2  (bases 1 to 5141)
  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: 
            "10.1073/pnas.1919600117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-02-11- 11"; volume: "117"; issue: "6"; pages: 
            "2978-2986"
COMMENT     Sequence Label: pTetOne-Six1-myc
FEATURES             Location/Qualifiers
     source          1..5141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..23
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    41..59
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    77..95
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    113..131
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1124..1213
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     polyA_signal    1360..1441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1474..1578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1579..1647,1648..2439)
                     /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     complement(1797..1816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2610..3198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3099..3118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3352..3369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3380..3461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3578..4324)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(4343..4853)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4406..4424)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    4873..4921
                     /note="synthetic polyadenylation signal"
     misc_feature    4935..5026
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4975..4994
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        join(5030..5141,1..256)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    5038..5056
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5074..5092
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5110..5128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
       61 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      121 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      181 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      241 ttcctaccct cgtaaagaat tcgccgccac catgtcgatg ctgccgtcgt ttggttttac
      301 gcaagagcaa gtggcgtgcg tgtgcgaagt tctgcagcaa ggagggaacc tggaacgcct
      361 gggcaggttc ttgtggtcgt tgcccgcctg cgatcacctg cacaagaacg agagcgtgct
      421 caaggccaag gcggtggtcg ccttccaccg cggcaacttc cgcgagctct acaagatact
      481 ggagagccac cagttctcgc ctcacaatca ccccaaactg cagcagctgt ggctgaaagc
      541 gcactacgtg gaggccgaga aacttcgcgg ccgacccctg ggtgccgtgg gcaaatatcg
      601 ggtgcgccga aaattcccgt tgccgcggac catctgggac ggcgaggaga ccagctactg
      661 ctttaaggag aagtctcggg gcgtgctgcg ggagtggtac gcgcacaacc cctacccctc
      721 accgagggag aaacgggagc tggccgaggc caccggcctc accaccaccc aggtcagcaa
      781 ctggtttaag aaccggaggc aaagagaccg ggccgccgag gccaaggaaa gggagaacac
      841 cgaaaacaat aactcctcct ccaacaagca gaatcaactc tctcctctgg aagggggcaa
      901 gccgctcatg tccagctcag aagaggagtt ctcacccccc caaagtccag accagaactc
      961 ggtccttctg ctccagagca atatgggcca cgccaggagc tcaaactatt ctcttccagg
     1021 cctcacagcc tcccagccca gccacggtct gcaagcccat cagcaccagc tccaggactc
     1081 tctgctgggc ccactcacct ccagtttggt ggacttgggt tccgaacaaa aactcatctc
     1141 agaagaggat ctggaacaaa aactcatctc agaagaggat ctggaacaaa aactcatctc
     1201 agaagaggat ctgtagaccg gtggatccag atctctgcag catatgtcta gaggatcata
     1261 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1321 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1381 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1441 ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1501 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1561 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1621 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1801 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2401 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2461 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3241 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3301 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3361 gtgagcgagg aagctcgggg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3421 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3481 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     3541 acaaatgtgg tatggctgat tatgatcctc tggagattta cccggggagc atgtcaaggt
     3601 caaaatcgtc aagagcgtca gcaggcagca tatcaaggtc aaagtcgtca agggcatcgg
     3661 ctgggagcat gtctaagtca aaatcgtcaa gggcgtcggt cggcccgccg ctttcgcact
     3721 ttagctgttt ctccaggcca catatgatta gttccaggcc gaaaaggaag gcaggttcgg
     3781 ctccctgccg gtcgaacagc tcaattgctt gtttcagaag tgggggcata gaatcggtgg
     3841 taggtgtctc tctttcctct tttgctactt gatgctcctg ttcctccaat acgcagccca
     3901 gtgtaaagtg gcccacggcg gacagagcgt acagtgcgtt ctccagggag aagccttgct
     3961 gacacaggaa cgcgagctga ttttccaggg tttcgtactg tttctctgtt gggcgggtgc
     4021 cgagatgcac tttagccccg tcgcgatgtg agaggagagc acagcggtat gacttggcgt
     4081 tgttccgcag aaagtcttgc catgactcgc cttccagggg gcaggagtgg gtatgatgcc
     4141 tgtccagcat ctcgattggc agggcatcga gcagggcccg cttgttcttc acgtgccagt
     4201 acagggtagg ctgctcaact cccagctttt gagcgagttt ccttgtcgtc aggccttcga
     4261 taccgactcc attgagtaat tccagagcag agtttatgac tttgctcttg tccagtctag
     4321 acatggtaat tcgatgatcc ccctggggag agaggtcggt gattcggtca acgagggagc
     4381 cgactgccga cgtgcgctcc ggaggcttgc agaatgcgga acaccgcgcg ggcaggaaca
     4441 gggcccacac taccgcccca caccccgcct cccgcaccgc cccttcccgg ccgctgctct
     4501 cggcgcgccc tgctgagcag ccgctattgg ccacagccca tcgcggtcgg cgcgctgcca
     4561 ttgctccctg gcgctgtccg tctgcgaggg tactagtgag acgtgcggct tccgtttgtc
     4621 acgtccggca cgccgcgaac cgcaaggaac cttcccgact taggggcgga gcaggaagcg
     4681 tcgccggggg gcccacaagg gtagcggcga agatccgggt gacgctgcga acggacgtga
     4741 agaatgtgcg agacccaggg tcggcgccgc tgcgtttccc ggaaccacgc ccagagcagc
     4801 cgcgtccctg cgcaaaccca gggctgcctt ggaaaaggcg caaccccaac cccgtggaat
     4861 tatcacctcg agaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4921 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     4981 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctctcg agtttactcc
     5041 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
     5101 agactttact ccctatcagt gatagagaac gtataaggag t
//
