LOCUS       sequence-12828-e        3596 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 3596)
  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 3596)
  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 R3L2 TetO(fl)-3
FEATURES             Location/Qualifiers
     source          1..3596
                     /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    complement(354..477)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    504..552
                     /note="synthetic polyadenylation signal"
     intron          700..1272
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1385..1409)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1406..1444)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(1477..1495,1496..1518,1519..1537,1538..1560,1561..1579,
                     1580..1602,1603..1621,1622..1644,1645..1663,1664..1686,
                     1687..1705,1706..1728,1729..1747)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    complement(1770..1803)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1825..1924)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2036..2055)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2047..2856
                     /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(2114..2133)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2946..3534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3435..3454
                     /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 tacttaagat ctccatagtg
      361 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
      421 atatattgat atttatatca ttttacgttt ctcgttcaac tttattatac atagttgctt
      481 aagatctctg aatgcggccg cttcacacaa aaaaccaaca cacagatgta atgaaaataa
      541 agatatttta ttgcggccat cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
      601 accccagagt cccgcctgca ggccggccta gtgcagggcg tcgacgaatt ctttgccaaa
      661 atgatgagac agcacaacaa ccagcacgtt gcccaggagc tgtaggaaag agaagaaggc
      721 atgaacatgg ttagcagagg ggcccggttt ggactcagag tattttatcc tcatctcaaa
      781 cagtgtatat cattgtaacc ataaagagaa aggcaggatg atgaccaggg tgtagttgtt
      841 tctaccaata agaatatttc cacgccagcc agaatttata tgcagaaata ttctacctta
      901 tcatttaatt ataacaattg ttctctaaaa ctgtgctgaa gtacaatata atataccctg
      961 attgccttga aaaaaaagtg attagagaaa gtacttacaa tctgacaaat aaacaaaagt
     1021 gaatttaaaa attcgttaca aatgcaagct aaagtttaac gaaaaagtta cagaaaatga
     1081 aaagaaaata agaggagaca atggttgtca acagagtaga aagtgaaaga aacaaaatta
     1141 tcatgagggt ccatggtgat acaagggaca tcttcccatt ctaaacaaca ccctgaaaac
     1201 tttgccccct ccatataaca tgaattttac aatagcgaaa aagaaagaac aatcaagggt
     1261 ccccaaactc accctgaagt tctcaggatc cccgggtacc gagctcgaat tcggggccgc
     1321 ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc
     1381 aggcgatctg acggttcact aaacgagctc tgcttatata ggcctcccac cgtacacgcc
     1441 tacctcgacc cgggtaccga gctcgacttt cacttttctc tatcactgat agggagtggt
     1501 aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt
     1561 tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc actgataggg
     1621 agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt
     1681 cacttttctc tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg
     1741 atagggagtg gtaaactcga taacctatga taacttcgta tagcatacat tatacgaagt
     1801 tatgtttaaa ctcgagatat ctagacccag ctttcttgta caaagttggc attataagaa
     1861 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1921 tttgccatcc agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     1981 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2041 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2101 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2161 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2221 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2281 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2341 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2401 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2461 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2521 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2581 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2641 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2701 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2761 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2821 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     2881 cattattcag attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3001 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
