LOCUS       sequence-9797-ex        5343 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 5343)
  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 5343)
  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-miR143Bul
FEATURES             Location/Qualifiers
     source          1..5343
                     /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    2060..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2125..2144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2503..2523
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2675..2692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2846..3434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2926..2945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3605..4396,4397..4465))
                     /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     4228..4247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4466..4570)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4597..5052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4684..4703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4894..4915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5183..5231
                     /note="synthetic polyadenylation signal"
     misc_feature    5245..5336
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5285..5304
                     /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 aattctagtg agctacagac gatcatctca gtgatcgagc tacagacgat
     1981 catctcagtg atcgagctac agacgatcat ctcaactagt actagagtcg gggcggccgg
     2041 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2101 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2161 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2221 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2281 aaggatctga acgatggagc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     2341 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     2401 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     2461 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     2521 tccacagcgg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     2581 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     2641 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     4621 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4681 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4741 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4801 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4861 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4921 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4981 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5041 cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5101 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
     5161 gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     5221 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
     5281 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg
     5341 ata
//
