LOCUS       sequence-245660-        5067 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 5067)
  AUTHORS   Heckler MM, Thakor H, Schafer CC, Riggins RB
  TITLE     ERK/MAPK regulates ERRgamma expression, transcriptional activity, 
            and receptor-mediated Tamoxifen resistance in ER+ breast cancer.
  JOURNAL   FEBS J. 2014 Mar 29. doi: 10.1111/febs.12797.
  PUBMED    24684682
REFERENCE   2  (bases 1 to 5067)
  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: "FEBS J. 
            2014 Mar 29. doi: 10.1111/febs.12797."
COMMENT     Sequence Label: 3xERRE/ERE-luciferase
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..197
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      48..183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      227..236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             233..1885
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(299..317)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1926..2047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1937..1956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1991..2010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2295..2312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2466..3054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3225..4016,4017..4085))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3848..3867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4086..4190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4217..4672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4514..4535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4803..4851
                     /note="synthetic polyadenylation signal"
     misc_feature    4865..4956
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4905..4924
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
       61 ctccgcccag ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag
      121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      181 gcctaggctt ttgcaaaaag cttggcattc cggtactgtt ggtaaagcca ccatggaaga
      241 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
      301 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      361 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
      421 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
      481 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
      541 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
      601 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
      661 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
      721 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
      781 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
      841 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
      901 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
      961 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     1021 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
     1081 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
     1141 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     1201 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     1261 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1321 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1381 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     1441 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     1501 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     1561 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     1621 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     1681 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     1741 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     1801 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     1861 gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc ttcgagcaga
     1921 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1981 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2041 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2101 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac
     2161 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     2221 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     2281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2881 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4201 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
     4681 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4741 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
     4801 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     4861 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     4921 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ataggtaccc ggacctcaag
     4981 gtcaccttga cctcgttcgg acctcaaggt caccttgacc tcgttcggac ctcaaggtca
     5041 ccttgacctg gatccagatc tgcgatc
//
