LOCUS       sequence-12118-e        7314 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 7314)
  AUTHORS   Levy C, Khaled M, Robinson KC, Veguilla RA, Chen PH, Yokoyama S, 
            Makino E, Lu J, Larue L, Beermann F, Chin L, Bosenberg M, Song JS, 
            Fisher DE
  TITLE     Lineage-specific transcriptional regulation of DICER by MITF in 
            melanocytes.
  JOURNAL   Cell. 2010 Jun 11. 141(6):994-1005.
  PUBMED    20550935
REFERENCE   2  (bases 1 to 7314)
  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: "Cell. 2010 
            Jun 11. 141(6):994-1005."
COMMENT     Sequence Label: pGL3-DICER-Prom downstream Ebox Mut
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2578..2587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2584..4236
                     /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(2650..2668)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4277..4398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4288..4307)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4342..4361
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4646..4663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4817..5405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4897..4916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5576..6367,6368..6436))
                     /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     6199..6218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6437..6541)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6568..7023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6655..6674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6865..6886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7154..7202
                     /note="synthetic polyadenylation signal"
     misc_feature    7216..7307
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7256..7275
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccaaaa tcagacaaac cggccaggcg cagtggctca tgcctgtaat cccaccactt
       61 tgggaagccg aggcaggtca ggagttcgag accagcctgg ccaacatggt gaaaccccgt
      121 ctctactgaa aatacaaaaa ttagccgggc gtggtggcac atgcctgtaa cccagctact
      181 tgggaggctg aggcagaaga atcgcttgaa cctaggaggc agaggttgca gtgagacgag
      241 attgcgccat tgcactccag cctgggcgac aagagtgaaa ctctgcctca aaaaagaaaa
      301 aaacaaatac ataaaattaa aaaaataaaa aaatcaggta tacctgctga tggaacaagc
      361 agcctggcta gtcttgacag attgcaagag ggcaaccaga ggggctgaag ttgtaccagc
      421 ccacaaattt ccactgccaa tagagtccag tgcctacccc catactgaga tgctgtcctt
      481 ctcataaagg cttatcaggg agtccaccga agagatatct acatttgggg ataagttaag
      541 acgatgttct ttaagccaga cacttattta acgcccaccg atattttaat tcctagctac
      601 aaagaaaacc ttatatgtat gactggggtc ttaaaggtat gacatgctca ccgtgcttga
      661 catataagaa actacttttt taaatacatg ggtctgccag gagcatccta cttcctaatt
      721 ttgcccataa atatgctact ttcaagtgaa accttaagcc aattcatagt aaatatatta
      781 tcaaaatttt agcacagtaa tgactgagca caccagcctc aaagccagac tacgtgggtt
      841 caaatctcag ttgttcactt actggctact tacttgttgg aagttattcc atttcttcgt
      901 ctgttaaaac aggaagcagg ctgggtacgg tggctcacgc ccgtaatccc agcactttgg
      961 gaggccaagg cggacggatg acaaggtcag aaatcgagac catcatggcc aacatggtga
     1021 aaccccgtct cttctaaaaa tacaaaaatt agctgggtgt ggtggcgtgc ccctgtaatc
     1081 ccagctactc gggaggctga ggcaggagaa tcacttgaac ctgggaggcg gagattgcag
     1141 taagcccagg tcgcaccact gcactccagc ctggagacag agtgagactc cgtctcaaaa
     1201 gaaaaaaaaa aaaaaaacag gaagcagttt cagcacctac ctcacaatgt gcacacagtt
     1261 aaagcaccta cctcacaggg tgcacacagt taaagcacct acctcacagg gtgcacctaa
     1321 ggatcacagg catgcaagcc ctagatcagc ctgcccaatg gcaatataat gcaggccaca
     1381 ggtgacattt aaaattctgt agaacccaca ctaaaaaata aacaaataaa acttttatcc
     1441 caatatatgg aaaaagttta tcacttcaac atgtaacaat tttttgaaca ttatttggta
     1501 aagaatgcat atatctcgat tccgactagc catactttaa gtgctcagga gcgaagtggg
     1561 gctggtggcc ccaaactgaa cagcacagcc caggcagcag gggcgcatag taggttctgc
     1621 tcacgcccat ggtctgaaag aaacacggtt gcagttgctg tgagcacacc tcacctggag
     1681 caaaccggcc tatcaggggg tccactgaag agatgtctac atttggggat aaattaagaa
     1741 gatgttcttt aagccagcca cttatttaat tcccaccgat attttaattc ctagcttcaa
     1801 gctgtggtgc gcggttccga cctatgcaaa tgaacagctt acaaacagag ggccaccatc
     1861 tatttctcat tatctgggga tcccggttgg ggtggggggc ggaagtgggt gtttgttatt
     1921 tccccaaatg cggaggcccc tgcaccacta acgcgaggaa acgcaggcga gtgggaaggt
     1981 actttaaaaa ggggagggag gtgctcagag ggaagctaag ctctccggga aacaattacg
     2041 gccacgttac ggtctgtggg tgccagaagg agcaacaatg gaggcagtcg aattccgaga
     2101 gctcagagag tctgccaaac ttagccggcc tcggcgcgcg ggttcccggg ggccccaaaa
     2161 actccgggca actgcggccc aacgccaagg tccagtccag gcgctgggcc gcaaacgcct
     2221 ctccggcctc cgcgcgcgcg cccgctgtct ccctgctcct gagtggctgg ggggccggga
     2281 ttaacctttc accgccaggc cctgtccaat cacaggctcg ctctcatgcc gccacggggc
     2341 gagcggagcg tgggggcgcg gaggggcggt gcgggagcca gtgggattct ccaagcggct
     2401 gtgcgccgtt gccgcgggcc gtgcgctggc tgccggccag ccgggcgctg cgcagtctcc
     2461 gcaggcgccg gcgggagggg tcgaggcgga ggcgcggcgc aggctgctgc aggcccaggt
     2521 gaatggagta acctgacagc ggggacgaaa gcttggcatt ccggtactgt tggtaaagcc
     2581 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
     2641 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
     2701 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
     2761 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
     2821 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     2881 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
     2941 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     3001 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
     3061 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     3121 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     3181 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     3241 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     3301 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     3361 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     3421 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     3481 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     3541 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     3601 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     3661 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     3721 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     3781 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     3841 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     3901 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     3961 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     4021 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     4081 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     4141 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     4201 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg
     4261 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4321 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4381 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4441 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4501 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     4561 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     4621 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4681 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4741 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4981 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5221 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5521 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5641 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5701 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5761 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6001 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6241 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6421 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6481 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6541 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     6601 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6661 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6721 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6781 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6841 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6901 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6961 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7021 tttcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7081 cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac
     7141 ttggagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     7201 tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc
     7261 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc gata
//
