LOCUS       V031872                 6478 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031872
VERSION     V031872
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..1914
                     /label="RNF175(NM_173662)"
                     /note="RNF175(NM_173662)"
                     /gene="RNF175"
     CDS             1921..2010
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2045..2269
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2315..2743
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2757..3086
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3153..3947
                     /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    4121..4242
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4291..4307)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4315..4331
                     /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(4339..4369)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4384..4405
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4693..5278)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5449..6309)
                     /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(6310..6414)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggccgcgg ggacggcggc gcggaaggca
      961 gcgccggtgc tggaggcccc cccgcagcag gagcagctct ctcatacaaa gctttctgca
     1021 gaagacacat ggaacctgca gcaggagagg atgtacaaga tgcaccgggg ccacgattcc
     1081 atgcacgtgg aaatgatctt gatcttcctc tgcgttctgg tcattgccca gatagtgctg
     1141 gttcagtgga gacagaggca tggccgatcc tacaatctgg tgaccttgtt gcagatgtgg
     1201 gttgtcccct tatatttcac gataaaatta tactggtggc ggtttctgtc tatgtggggg
     1261 atgttctccg ttattaccag ttacatcctc ttcagagcta cccgaaaacc cctctcagga
     1321 aggacaccac gattggtcta caaatggttt cttttgatct acaaactcag ctatgcattt
     1381 ggtgttgtgg gttacttggc gatcatgttt acaatgtgtg gattcaatct gtttttcaaa
     1441 atcaaagcta gagattccat ggattttggc attgtgtctt tgttctacgg cctctactat
     1501 ggagtaatgg ggagagactt tgccgagatc tgctcagact acatggcttc cactataggg
     1561 ttctacagtg tcagccggtt gcctacaagg agcttatcgg acaatatctg tgcagtctgt
     1621 gggcagaaga tcattgtgga gcttgatgaa gaagggctca ttgaaaacac ctaccagctt
     1681 tcctgtaatc atgtctttca tgaattctgc atccgaggtt ggtgtatcgt tgggaaaaag
     1741 cagacttgcc cttactgcaa agagaaagtt gatttgaaga ggatgatcag taatccctgg
     1801 gagcgcacac attttctgta tggacaaatc ctggattggc ttcgttattt ggtggcctgg
     1861 caacctgtgg tgataggaat agttcaaggc attatctatt cactagggct ggaactcgag
     1921 gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga agaagattta
     1981 gagcaaaagt taatttccga ggaggactta tgagggcccg tttaaacccg ctgatcagcc
     2041 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2101 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2161 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2221 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2281 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2341 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2401 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2461 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2521 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2581 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2641 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2701 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2761 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2821 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2881 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2941 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3001 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3061 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3121 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3181 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3241 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3301 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3361 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3421 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3481 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3541 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3601 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3661 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3721 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3781 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3841 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3901 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3961 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4021 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4081 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4141 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4201 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4261 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4321 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4381 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4441 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4501 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5221 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6421 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
