LOCUS       sequence-9795-ex        5346 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 5346)
  AUTHORS   Hwang HW, Wentzel EA, Mendell JT
  TITLE     Cell-cell contact globally activates microRNA biogenesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7016-21. doi: 
            10.1073/pnas.0811523106.
  PUBMED    19359480
REFERENCE   2  (bases 1 to 5346)
  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"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2009-04-28- 28"; 
            volume: "106"; issue: "17"; pages: "7016-21"
COMMENT     Sequence Label: pGL3con-miR15aBul
FEATURES             Location/Qualifiers
     source          1..5346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..1932
                     /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(346..364)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2063..2184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2074..2093)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2128..2147
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2506..2526
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2678..2695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2849..3437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2929..2948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3608..4399,4400..4468))
                     /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     4231..4250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4469..4573)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4600..5055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4687..4706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4897..4918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5186..5234
                     /note="synthetic polyadenylation signal"
     misc_feature    5248..5339
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5288..5307
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      301 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      361 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
      421 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
      481 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
      541 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
      601 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
      661 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
      721 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
      781 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
      841 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
      901 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
      961 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1021 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1081 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1141 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1201 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1261 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1321 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1381 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     1441 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     1501 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     1561 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     1621 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     1681 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     1741 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     1801 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     1861 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     1921 atcgccgtgt aattctagtc acaaaccata tactgctgct agtgatccac aaaccatata
     1981 ctgctgctag tgatccacaa accatatact gctgctaact agtactagag tcggggcggc
     2041 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2101 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2161 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2221 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2281 gataaggatc tgaacgatgg agcggagaat gggcggaact gggcggagtt aggggcggga
     2341 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     2401 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     2461 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     2521 cattccacag cggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     2581 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     2641 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2701 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2761 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2821 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2881 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2941 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3001 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3061 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3541 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3601 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3661 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3721 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3781 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3841 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3901 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3961 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4021 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4081 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4141 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4201 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4261 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4321 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4381 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4441 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4501 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4561 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4621 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4681 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4741 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4801 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4861 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4921 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4981 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5041 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5101 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     5161 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5221 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     5281 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta
     5341 tcgata
//
