LOCUS       V047170                 8394 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047170
VERSION     V047170
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3074
                     /label="Dgcr8(NM_033324)"
                     /note="Dgcr8(NM_033324)"
                     /gene="Dgcr8"
     CDS             3081..3146
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3170..3758
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3824..3872
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4074..4502
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4516..4845
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4912..5706
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5882..6003
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6052..6068)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6076..6092
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6100..6130)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6145..6166
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6454..7042)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7213..8073)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8074..8178)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga gacatatgag agtccctctc
      781 ctctcccgcg tgagcccgca ggagaagcga tgatggagaa ccgagcttgc cccttccaag
      841 tgctgcccca tgaacagtct ccaccacctc ccctgcaaac gtccagtgat gcagaggtaa
      901 tggacgttgg ctctggtggt gatggacagt ccgaacctcc tgccgacgac ccattcaact
      961 tctacggagc ttctcttctc tccaaaggat ccttctctaa gggccgcctc ctcatagacc
     1021 cgaactgtag tggccacagc ccgcgcactg cccggcacgc acctgcggtc cggaagttct
     1081 cccctgacct taagttgctt aaggatgtaa agattagcgt gagctttact gagagctgca
     1141 ggagtaagga caggaaggtg ctgtacacag gagtagaacg cagcactcgg cctgagtgtg
     1201 gccagctcct tagtcctgtc agtggggacg tgcatgcttg tccctttggc gggagtgttg
     1261 gtaatggggt aggcctaggg ggtgagagtg cagataagaa ggatgaggaa aatgagctgg
     1321 atcaggaaaa gagagtggag tatgcagtgc tcgatgagtt agaagatttt actgacaatt
     1381 tggagctaga tgaagaagga acaggcgggt tcacggctaa agcaatcgtt caaagagaca
     1441 gagtggatga agaggccttg aatttctcct atgaggatga ctttgacaat gatgtggacg
     1501 ctttactaga agaaggtctc tgtgctccca agaagaggcg aatggaggaa aaatatggcg
     1561 gagacagtga tcatccatct gatggagaga caagtgtaca gccaatgatg accaagatta
     1621 aaacagtgct caaaagtcgt ggccgtccac ctacagagcc attgcctgat ggatggatca
     1681 tgacttttca taattctgga gtccctgtat acctgcacag agagtctcga gtggtcactt
     1741 ggtccagacc ctacttcttg ggaacaggaa gcatacggaa acatgatcct cctctaagca
     1801 gtatcccctg cctacattat aagaaaatga aggacaatga ggaacgagaa caaaactgtg
     1861 atcttgcccc cagtggagag gtgtcacctg tcaagccctt gggtcggtct gcagagttgg
     1921 atttccctct ggaagagcct gactccatgg gtggagactc agggtccatg gatgagaagg
     1981 acccattggg ggctgaggca gccgctggag ccctgggaca agtgaaggct aaagttgagg
     2041 tgtgcaaaga tgaatcagtt gatctggagg aatttcgtaa ttaccttgag aagcgttttg
     2101 actttgaaca agtaactgtg aaaaaattca ggacttgggc tgagcggcgt cagttcaacc
     2161 gtgagatgaa gcggaagcag gccgagtcag agaggcccat cctgccagcc aaccagaagc
     2221 tgatcactct atctgtacaa gatgcaccca caaagaaaga gtttgtcatc aatcccaatg
     2281 ggaagtctga ggtttgcatc ctgcacgaat acatgcagcg tgtcctcaag gtccgccctg
     2341 tttataattt ctttgaatgt gagaatccaa gtgagccttt tggtgcctcc gtgaccattg
     2401 atggtgtgac ttacggatct ggaactgcaa gcagcaaaaa acttgcgaag aataaagctg
     2461 cccgagccac cctggaaatt ctcatccctg actttgttaa acagacctct gaggagaagc
     2521 ctaaagacag tgaagaactg gagtatttta accacatcag tattgaggat tcacgagtct
     2581 atgagctgac aagcaaggct gggctgttgt ctccatatca gatcctccat gagtgcctta
     2641 aaagaaacca tggaatgggt gacacatcca tcaagtttga agtggttcct gggaaaaacc
     2701 agaagagtga atatgttatg gcatgcggca aacacacagt gcgcgggtgg tgtaagaata
     2761 aacgagttgg gaaacaatta gcatctcaga aaatccttca gctactgcac ccacatgtca
     2821 agaactgggg ttccttacta cgcatgtatg gtcgtgagag cagcaaaatg gtcaagcagg
     2881 agacctctga caagagtgtg atagagctac agcagtatgc caagaagaac aggcccaacc
     2941 ttcacatcct gagcaagcta caagaagaga tgaagaggct ggctgcagag cgggaggaga
     3001 ctcggaagaa acccaagatg tcaattgtag catctgccca gcctggtggt gagcccttgt
     3061 gcacagtcga tgtaggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3121 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3181 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3241 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3301 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3361 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3421 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3481 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3541 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3601 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3661 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3721 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3781 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3841 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3901 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3961 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4021 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4081 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4141 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4201 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4261 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4321 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4381 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4441 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4501 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4561 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4621 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4681 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4741 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4801 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4861 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4921 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4981 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5041 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5101 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5161 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5221 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5281 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5341 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5401 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5461 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5521 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5581 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5641 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5701 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5881 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6001 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8221 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8281 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8341 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
