LOCUS       sequence-322827-        5911 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 5911)
  AUTHORS   O'Donnell KA, Wentzel EA, Zeller KI, Dang CV, Mendell JT
  TITLE     c-Myc-regulated microRNAs modulate E2F1 expression.
  JOURNAL   Nature. 2005 Jun 9. 435(7043):839-43.
  PUBMED    15944709
REFERENCE   2  (bases 1 to 5911)
  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: "Nature. 
            2005 Jun 9. 435(7043):839-43."
COMMENT     Sequence Label: pGL3-Control-E2F-3'UTR-WT
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        795..991
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      842..977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1021..1030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1027..2679
                     /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(1093..1111)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3375..3496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3386..3405)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3440..3459
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3818..3838
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3990..4007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4161..4749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4241..4260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4920..5711,5712..5780))
                     /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     5543..5562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgataggt accgagctct tacgcgtgct agcccgggct
      781 cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      841 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt
      901 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
      961 aggctttttt ggaggcctag gcttttgcaa aaagcttggc attccggtac tgttggtaaa
     1021 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa
     1081 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
     1141 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa
     1201 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
     1261 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
     1321 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
     1381 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac
     1441 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac
     1501 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
     1561 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc
     1621 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga
     1681 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     1741 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     1801 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag
     1861 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc
     1921 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc
     1981 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa
     2041 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     2101 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     2161 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg
     2221 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     2281 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg
     2341 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg
     2401 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt
     2461 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     2521 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     2581 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     2641 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagatgtg tgcatgagtc
     2701 catgtgtgcg cgtggggggg ctctaactgc actttcggcc cttttgctct gggggtccca
     2761 caaggcccag ggcagtgcct gctcccagaa tctggtgctc tgaccaggcc aggtggggag
     2821 gctttggctg gctgggcgtg taggacggtg agagcacttc tgtcttaaag gttttttctg
     2881 attgaagctt taatggagcg ttatttattt atcgaggcct ctttggtgag cctggggaat
     2941 cagcaaaggg gaggaggggt gtggggttga taccccaact ccctctaccc ttgagcaagg
     3001 gcaggggtcc ctgagctgtt cttctgcccc atactgaagg aactgaggcc tgggtgattt
     3061 atttattggg aaagtgaggg agggagacag actgactgac agccatgggt ggtcagatgg
     3121 tggggtgggc cctctccagg gggccagttc agggccccag ctgcccccca ggatggatat
     3181 gagatgggag aggtgagtgg gggaccttca ctgatgtggg caggaggggt ggtgaaggcc
     3241 tcccccagcc cagaccctgt ggtccctcct gcagtgtctg aagcgcctgc ctccccactg
     3301 ctctgcccca ccctccaatc tgcactttga tttgctctag agtcggggcg gccggccgct
     3361 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3421 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3481 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3541 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3601 tctgaacgat ggagcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     3661 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     3721 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
     3781 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
     3841 agcggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     3901 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     3961 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct g
//
