LOCUS       sequence-478992-        7791 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  AUTHORS   Yeom KH, Lee Y, Han J, Suh MR, Kim VN
  TITLE     Characterization of DGCR8/Pasha, the essential cofactor for Drosha 
            in primary miRNA processing.
  JOURNAL   Nucleic Acids Res. 2006;34(16):4622-9. Epub 2006 Sep 8.
  PUBMED    16963499
REFERENCE   2  (bases 1 to 7791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2006"; volume: "34(16)"; pages: "4622-9. Epub 
            2006 Sep 8"
COMMENT     Sequence Label: pcDNA3-V5-DGCR8
FEATURES             Location/Qualifiers
     source          1..7791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             898..939
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        complement(3344..3362)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3345..3362)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3382..3399)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3388..3612
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3658..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3745..3764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3955..3976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4095..4115)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4100..4429
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4280..4415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4342..4361
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4496..5290
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4550..4569)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5160..5179
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5464..5585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5501..5520)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5555..5574
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5634..5650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5634..5650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5647..5669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5658..5674
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5682..5688,5689..5706,5707..5712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5727..5748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5865..5882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6036..6624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6116..6135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6795..7586,7587..7655))
                     /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     7418..7437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7656..7760)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgggt
      901 aagcctatcc ctaaccctct cctcggtctc gattctacgg gatccactag taacggccgc
      961 cagtgtgctg gaattcgatt gaatctgatg gagacagatg agagcccctc tccgctcccg
     1021 tgtgggcccg caggagaagc ggtgatggag agccgagctc gccccttcca agcgctgccc
     1081 cgtgagcagt ctccaccacc tcccctgcaa acgtccagtg gtgcagaggt aatggacgtt
     1141 ggctctggtg gtgatggaca gtccgaactc cctgctgagg accccttcaa cttctacgga
     1201 gcttctcttc tctccaaagg atccttctct aagggccgcc tcctcataga cccgaactgt
     1261 agtggccaca gcccgcgcac cgcccggcac gcacctgcgg tccggaagtt ctcccctgac
     1321 cttaagttgc ttaaggatgt aaagattagc gtgagcttta ccgagagctg caggagtaag
     1381 gacaggaagg tgctgtacac aggagcagag cgcgacgtgc gggcggagtg cggtctgctc
     1441 cttagccctg tcagtgggga cgtgcatgct tgtccctttg gcgggagtgt tggtgacggg
     1501 gtaggcatag ggggtgagag tgctgataag aaggatgagg agaatgagct ggatcaggaa
     1561 aagagagtgg agtatgcagt gctcgatgag ttagaagatt ttactgacaa tttggagcta
     1621 gatgaagaag gagcaggcgg gttcacggct aaagcaatcg ttcagagaga cagagtggat
     1681 gaagaggcct tgaatttccc ctacgaggat gactttgaca acgatgtgga tgctctgctg
     1741 gaagaaggcc tttgtgcccc caaaaagagg cgaacagagg aaaaatatgg cggagacagc
     1801 gaccatccgt ccgatggaga gacaagtgtg cagccgatga tgaccaagat taaaacagtg
     1861 ctcaaaagtc gtggccgccc acctacagag ccgctgcccg acgggtggat catgacattc
     1921 cataactctg gagtcccggt gtacctacac agagagtctc gggtggtcac ctggtccagg
     1981 ccatacttct tgggaacggg aagcatacgg aaacacgacc ctcctctgag tagcatccct
     2041 tgtctgcatt ataagaaaat gaaggacaac gaggaacggg agcaaagcag tgacctcacc
     2101 cctagtgggg atgtgtcccc cgtcaagccc ctgagccgat ctgcagagct ggagtttccc
     2161 ctggatgagc ctgactctat gggtgctgac ccggggcccc cggacgagaa agacccacta
     2221 ggggctgagg cagcccctgg ggccctgggg caggtgaagg ccaaagtcga ggtgtgcaaa
     2281 gatgaatccg ttgatctcga ggaatttcga agctacctgg agaagcgttt tgactttgag
     2341 caagttactg tgaaaaaatt caggacttgg gctgagcggc ggcaattcaa tcgggaaatg
     2401 aagcggaagc aggcggagtc cgagaggccc atcttgccag ccaatcagaa gctcattact
     2461 ttatcagtgc aagatgcacc cacaaagaaa gagtttgtta ttaaccccaa cgggaaatcc
     2521 gaggtctgca tcctgcacga gtacatgcag cgtgtcctca aggtccgccc tgtctataat
     2581 ttctttgaat gtgagaaccc aagtgagcct tttggtgcct cggtgaccat tgatggtgtg
     2641 acttacggat ctggaactgc aagcagcaaa aaacttgcga agaataaagc tgcccgagct
     2701 acactggaaa tcctcatccc tgactttgtt aaacagacct ctgaagagaa gcccaaagac
     2761 agtgaagaac tcgagtattt taaccacatc agcatcgagg actcgcgggt ctacgagctg
     2821 accagcaagg ctgggctgtt gtctccatat cagatcctcc acgagtgcct taaaagaaac
     2881 catgggatgg gtgacacgtc tatcaagttt gaagtggttc ctgggaaaaa ccagaagagt
     2941 gaatacgtca tggcgtgtgg caagcacaca gtgcgcgggt ggtgtaagaa caagagagtt
     3001 ggaaagcagt tagcctcaca gaagatcctt cagctgctgc acccacatgt caagaactgg
     3061 gggtccttac tgcgcatgta tggccgtgag agcagcaaga tggtcaagca ggagacatcg
     3121 gacaagagtg tgattgagct gcagcagtat gccaagaaga acaagcccaa cctgcacatc
     3181 ctcagcaagc tccaagagga gatgaagagg ctagctgagg aaagggagga gactcgaaag
     3241 aagcccaaga tgtccattgt ggcgtccgcc cagcctggcg gtgagcccct gtgcaccgtg
     3301 gacgtgtgag cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     3361 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3421 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3481 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3541 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3601 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3841 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4081 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4441 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4501 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4561 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4621 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4681 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4741 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4801 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4861 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4921 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4981 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5041 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5101 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5161 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5221 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5281 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5341 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5401 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5461 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5521 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5581 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5641 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5701 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5761 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5821 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5881 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6721 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6781 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6841 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6901 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6961 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7021 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7081 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7141 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7201 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7261 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7321 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7381 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7441 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7501 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7561 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7621 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7681 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7741 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
