LOCUS       sequence-246573-        5846 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 5846)
  AUTHORS   Neuman E, Flemington EK, Sellers WR, Kaelin WG
  TITLE     Transcription of the E2F-1 gene is rendered cell cycle dependent by 
            E2F DNA-binding sites within its promoter.
  JOURNAL   Mol Cell Biol. 1994 Oct . 14(10):6607-15.
  PUBMED    7935380
REFERENCE   2  (bases 1 to 5846)
  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: "Mol Cell 
            Biol. 1994 Oct . 14(10):6607-15."
COMMENT     Sequence Label: pGL2-AN
FEATURES             Location/Qualifiers
     source          1..5846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             324..1976
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(390..408)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2217..2282
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2412..2432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2857..2991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3133..3150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3304..3892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3384..3403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4063..4854,4855..4923))
                     /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     4686..4705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4924..5028)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5055..5510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5142..5161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5352..5373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5700..5821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cccgggaggt accatccgga caaagcctgc gcgcgccccg ccccgccatt ggccgtaccg
       61 ccccgcgccg ccgccccatc tcgcccctcg ccgccgggtc cggcgcgtta aagccaatag
      121 gaaccgccgc cgttgttccc gtcacggccg gggcagccaa ttgtggcggc gctcggcggc
      181 tcgtggctct ttcgcggcaa aaaggatttg gcgcgtaaaa gtggccggga ctttgcaggc
      241 agcggcggcc gggggcggag cgggatcgag ccctcgatga tatcagatct aagtaagctt
      301 ggcattccgg tactgttggt aaaatggaag acgccaaaaa cataaagaaa ggcccggcgc
      361 cattctatcc tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat
      421 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt
      481 acgcggaata cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga
      541 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt
      601 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg
      661 aattgctcaa cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt
      721 tgcaaaaaat tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg
      781 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac
      841 ctcccggttt taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg
      901 cactgataat gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata
      961 gaactgcctg cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc
     1021 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac
     1081 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt
     1141 ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt
     1201 cattcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg
     1261 cttctggggg cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc
     1321 ttccagggat acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac
     1381 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg
     1441 ttgtggatct ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca
     1501 gaggacctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg
     1561 acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct
     1621 tcatagttga ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg
     1681 aattggaatc gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc
     1741 ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga
     1801 tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc
     1861 gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa
     1921 gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaaaatg
     1981 taactgtatt cagcgatgac gaaattctta gctattgtaa tactgcgatg agtggcaggg
     2041 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     2101 aataagtgat aataagcgga tgaatggcag aaattcgccg gatctttgtg aaggaacctt
     2161 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     2221 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     2281 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     2341 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     2401 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     2461 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     2521 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa
     2581 cctttataag taggcataac agttataatc ataacatact gttttttctt actccacaca
     2641 ggcatagagt gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa
     2701 tttgtaaagg ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc
     2761 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2821 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2881 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2941 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgtcgaccg
     3001 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     3061 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3121 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5041 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5101 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5401 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc
     5521 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5581 gcccaagcta ccatgataag taagtaatat taaggtacgt ggaggtttta cttgctttaa
     5641 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5821 atggtactgt aactgagcta acataa
//
