LOCUS       sequence-142926-        6127 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 6127)
  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 6127)
  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-PTEN-3'UTR-Wt
FEATURES             Location/Qualifiers
     source          1..6127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2571..2619
                     /note="synthetic polyadenylation signal"
     misc_feature    2633..2724
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2673..2692
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2779..2975
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2826..2961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3005..3014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3011..4663
                     /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(3077..3095)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5575..5696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5586..5605)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5640..5659
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6018..6038
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg
     2461 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta
     2521 ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat
     2581 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg
     2641 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     2701 gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcttacgcg tgctagcccg
     2761 ggctcgagat ctgcgatctg catctcaatt agtcagcaac catagtcccg cccctaactc
     2821 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat cgctgactaa
     2881 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2941 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tggcattccg gtactgttgg
     3001 taaagccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     3061 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     3121 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     3181 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     3241 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     3301 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     3361 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     3421 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     3481 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     3541 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     3601 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     3661 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     3721 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     3781 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct
     3841 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     3901 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     3961 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     4021 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     4081 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     4141 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     4201 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     4261 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
     4321 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     4381 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
     4441 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     4501 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     4561 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     4621 gatcctcata aaggccaaga agggcggaaa gatcgccgtg taattctaga ggctaaagag
     4681 ctttgtgata tactggttca catcctaccc ctttgcactt gtggcaacag ataagtttgc
     4741 agttggctaa gagaggtttc cgaagggttt tgctacattc taatgcatgt attcgggtta
     4801 ggggaatgga gggaatgctc agaaaggaaa taattttatg ctggactctg gaccatatac
     4861 catctccagc tatttacaca cacctttctt tagcatgcta cagttattaa tctggacatt
     4921 cgaggaattg gccgctgtca ctgcttgttg tttgcgcatt tttttttaaa gcatattggt
     4981 gctagaaaag gcagctaaag gaagtgaatc tgtattgggg tacaggaatg aaccttctgc
     5041 aacatcttaa gatccacaaa tgaagggata taaaaataat gtcataggta agaaacacag
     5101 caacaatgac ttaaccatat aaatgtggag gctatcaaca aagaatgggc ttgaaacatt
     5161 ataaaaattg acaatgattt attaaatatg ttttctcaat tgtaacgact tctccatctc
     5221 ctgtgtaatc aaggccagtg ctaaaattca gatgctgtta gtacctacat cagtcaacaa
     5281 cttacactta ttttactagt tttcaatcat aatacctgct gtggatgctt catgtgctgc
     5341 ctgcaagctt cttttttctc attaaatata aaatattttg taatgctgca cagaaatttt
     5401 caatttgaga ttctacagta agcgtttttt ttctttgaag atttatgatg cacttattca
     5461 atagctgtca gccgttccac ccttttgacc ttacacattc tattacaatg aattttgcag
     5521 ttttgcacat ttttttctag agtcggggcg gccggccgct tcgagcagac atgataagat
     5581 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     5641 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     5701 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     5761 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tctgaacgat ggagcggaga
     5821 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5881 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5941 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6001 gcctggggac tttccacacc ctaactgaca cacattccac agcggatccg tcgaccgatg
     6061 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     6121 gccgcac
//
