LOCUS       V042332                 8644 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042332
VERSION     V042332
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"
     CDS             706..3303
                     /label="RNF216(NM_001377156)"
                     /note="RNF216(NM_001377156)"
                     /gene="RNF216"
     CDS             3310..3336
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3340..3366
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3370..3396
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3420..4008
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4074..4122
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4324..4752
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4766..5095
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5162..5956
                     /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    6132..6253
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6302..6318)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6326..6342
                     /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(6350..6380)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6395..6416
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6704..7292)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7463..8323)
                     /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(8324..8428)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga agagggaaac
      721 aacaatgaag aggtaattca cttgaacaac tttcactgcc atcggggaca agagtggatc
      781 aatctccgag atgggcccat caccatatct gactcctcag atgaggaaag gattccaatg
      841 ctggtcaccc cagctcctca gcagcatgaa gaagaggacc tggatgatga tgtcatcctg
      901 acagaagatg attctgagga tgactacggt gaatttctgg atcttgggcc tcctggaatc
      961 tctgaattca ctaagccaag tggccaaaca gaaagagaac ccaagcctgg accgagtcat
     1021 aaccaagcag caaatgacat tgtcaacccc agatcagagc agaaagtcat catcttggaa
     1081 gaaggtagcc ttctttacac agaaagcgat cctttggaaa ctcagaacca gtcatccgaa
     1141 gactcagaga cagagctgtt atcaaatcta ggagagtcag ctgctctagc agatgatcag
     1201 gccatcgaag aagactgctg gttagatcat ccttacttcc agtctctgaa ccaacagccc
     1261 cgtgaaataa caaaccaggt cgttcctcag gaacggcagc ctgaagcaga actgggccgc
     1321 ttgttgtttc agcatgaatt cccagggccc gcttttccaa ggccggaacc ccagcaaggt
     1381 gggatttcag gcccctcttc tcctcagcct gcccatcctc taggagagtt tgaagaccag
     1441 cagttagcaa gtgatgatga agagccaggt ccagcctttc caatgcaaga atctcaagag
     1501 cccaatttgg aaaacatttg ggggcaagaa gctgcagagg tagatcaaga gctcgttgaa
     1561 ctactagtga aagaaacgga agcaagattt ccagatgtag caaatgggtt tattgaggaa
     1621 ataattcatt ttaagaatta ttatgatctg aatgtacttt gtaattttct tctggaaaac
     1681 ccagattatc caaagagaga agacagaatc attataaatc ccagtagcag tctgctggcc
     1741 agccaagatg agacaaagtt gcctaaaata gacttttttg actattctaa attgacccct
     1801 cttgaccagc gctgcttcat ccaagctgct gacctcctca tggccgactt caaagtgctc
     1861 agtagtcagg acatcaagtg ggccctgcac gagctcaaag gacactatgc aatcacccga
     1921 aaggccttgt ctgatgccat taaaaaatgg caggagctgt caccagaaac cagtggaaaa
     1981 aggaagaaga gaaaacaaat gaaccagtat tcttacattg atttcaagtt tgaacaaggt
     2041 gacataaaaa tagaaaagag gatgttcttt cttgaaaata agcgacgaca ttgtaggtcc
     2101 tatgaccgac gtgctctcct tccagctgtg caacaagagc aggagttcta tgagcagaaa
     2161 atcaaagaga tggcagagca tgaagacttt ttgcttgccc tacagatgaa tgaagaacag
     2221 tatcaaaagg atggccagct gattgagtgt cgctgctgct atggggaatt tccattcgag
     2281 gagctgacgc agtgcgcaga tgctcacttg ttctgcaaag agtgtctcat cagatatgcc
     2341 caagaggcag tctttggatc tggaaagttg gagctcagct gcatggaagg cagctgcacg
     2401 tgttcgttcc caaccagtga gctggagaag gtgctccccc agaccatcct gtataagtac
     2461 tatgagcgaa aagccgagga ggaggttgcg gcagcctacg ccgacgagct tgtcaggtgc
     2521 ccgtcctgta gctttccggc tctgttggac agtgatgtga agaggttcag ctgtcctaat
     2581 cctcactgcc gaaaggaaac ctgtaggaag tgtcagggac tctggaaaga acataatggc
     2641 ctcacctgtg aagagctggc tgaaaaagac gacatcaagt accgtacctc tattgaagaa
     2701 aaaatgactg ctgcccgcat tagaaaatgc cacaagtgtg ggactggcct catcaaatct
     2761 gaaggctgca accgcatgtc ttgccgctgt ggtgcccaga tgtgctacct ctgtcgagtt
     2821 tctattaatg gatatgacca tttctgccaa catccccgct caccaggagc cccttgccag
     2881 gagtgttcaa gatgctctct ctggaccgat cccactgaag atgatgagaa gcttattgag
     2941 gaaatccaga aggaggctga agaggaacag aaaagaaaga atggagagaa caccttcaaa
     3001 cgcattggac ccccgctgga gaagcctgtg gagaaggtgc agagggtgga ggccctcccg
     3061 aggcccgttc cgcagaacct gccacagcca cagatgccac cctatgcctt cgcgcaccca
     3121 cccttccccc tgcctcccgt gcggcctgtg ttcaacaact tcccactcaa catggggcct
     3181 atcccagccc cgtacgtgcc ccctctgccc aacgtgcggg tcaactatga cttcggtccc
     3241 atccacatgc ccctggagca caacctgccc atgcactttg gcccccagcc gcggcatcgc
     3301 ttcctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     3361 tatgctagct atccttatga tgtcccagac tacgcatagt gagggcccga tatctcgaca
     3421 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3481 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3541 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3601 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3661 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3721 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3781 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3841 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3901 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3961 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     4021 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4081 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4141 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4201 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4261 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     4321 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4381 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4441 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     4501 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4561 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4621 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4681 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4741 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4921 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4981 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     5041 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5101 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5161 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5221 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5281 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5341 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5401 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5461 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5521 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5581 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5641 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5701 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5761 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5821 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5881 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5941 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     6001 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6061 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6121 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6241 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6301 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6361 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6421 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6541 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6601 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6661 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6721 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6781 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6841 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6901 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6961 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7021 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7081 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7141 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7201 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7261 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7321 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7381 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7441 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8341 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8401 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8461 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     8521 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     8581 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     8641 tagg
//
