LOCUS       sequence-12762-e        3420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3420)
  AUTHORS   Hsiao EC, Nguyen TD, Ng JK, Scott MJ, Chang WC, Zahed H, Conklin BR
  TITLE     Constitutive Gs activation using a single-construct 
            tetracycline-inducible expression system in embryonic stem cells and
            mice.
  JOURNAL   Stem Cell Res Ther. 2011 Mar 4. 2(2):11.
  PUBMED    21375737
REFERENCE   2  (bases 1 to 3420)
  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: "Stem Cell 
            Res Ther. 2011 Mar 4. 2(2):11."
COMMENT     Sequence Label: pEnt L1L3 rtTA-3
FEATURES             Location/Qualifiers
     source          1..3420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(490..523)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(607..1224,1225..1230,1231..1266,1267..1269,1270..1305,
                     1306..1308,1309..1344,1345..1353)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(1363..1379)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1363..1379)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1389..1407)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1390..1407)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1427..1444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1433..1640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1655..1750)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1860..1879)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1871..2680
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1938..1957)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2770..3358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3259..3278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 cgtttaaaca taacttcgta tagcatacat tatacgaagt tatctcgagc ctcaggaaac
      541 agggtacccg gggatccaca tatgttaatt aagactcctg caggattaag cccgggcgaa
      601 ttcaccatgt ctagactgga caagagcaaa gtcataaacg gcgctctgga attactcaat
      661 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
      721 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
      781 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
      841 ctgcggaaca acgccaagtc attccgctgt gctctcctct cacatcgcga cggggctaaa
      901 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
      961 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     1021 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     1081 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
     1141 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     1201 aaacagctaa agtgcgaaag cggcgggccg gccgacgccc ttgacgattt tgacttagac
     1261 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     1321 gacgattttg accttgacat gctccccggg taactaagta aggatccact agttctagag
     1381 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     1441 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1501 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1561 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1621 aatagcaggc atgctgggga gcggccgcag atctcaactt tattatacaa agttggcatt
     1681 ataaaaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1741 atcattattt gatatcctgc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1801 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1861 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1921 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1981 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2041 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2101 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2161 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2221 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2281 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2341 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2401 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2461 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2521 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2581 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2641 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2701 gtaacattat tcagattggg ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
