LOCUS       V020055                 8609 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020055
VERSION     V020055
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             896..1603
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1625..4129
                     /label="TRIM28(NM_005762)"
                     /note="TRIM28(NM_005762)"
                     /gene="TRIM28"
     polyA_signal    4176..4400
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4446..4874
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4888..5217
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5284..6078
                     /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    6252..6373
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6422..6438)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6446..6462
                     /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(6470..6500)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6515..6536
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6824..7409)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7580..8440)
                     /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(8441..8545)
                     /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 ccaccatggt
      901 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
      961 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1021 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1081 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1141 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1201 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1261 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1321 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1381 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1441 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1501 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1561 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtggatcg gaggaggggg
     1621 aagcatggcg gcctccgcgg cggcagcctc ggcagcagcg gcctcggccg cctctggcag
     1681 cccgggcccg ggcgagggct ccgctggcgg cgaaaagcgc tccaccgccc cttcggccgc
     1741 agcctcggcc tctgcctcag ccgcggcgtc gtcgcccgcg gggggcggcg ccgaggcgct
     1801 ggagctgctg gagcactgcg gcgtgtgcag agagcgcctg cgacccgaga gggagccccg
     1861 cctgctgccc tgtttgcact cggcctgtag tgcctgctta gggcccgcgg cccccgccgc
     1921 cgccaacagc tcgggggacg gcggggcggc gggcgacggc accgtggtgg actgtcccgt
     1981 gtgcaagcaa cagtgcttct ccaaagacat cgtggagaat tatttcatgc gtgatagtgg
     2041 cagcaaggct gccaccgacg cccaggatgc gaaccagtgc tgcactagct gtgaggataa
     2101 tgccccagcc accagctact gtgtggagtg ctcggagcct ctgtgtgaga cctgtgtaga
     2161 ggcgcaccag cgggtgaagt acaccaagga ccatactgtg cgctctactg ggccagccaa
     2221 gtctcgggat ggtgaacgta ctgtctattg caacgtacac aagcatgaac cccttgtgct
     2281 gttttgtgag agctgtgata ctctcacctg ccgagactgc cagctcaatg cccacaagga
     2341 ccaccagtac cagttcttag aggatgcagt gaggaaccag cgcaagctcc tggcctcact
     2401 ggtgaagcgc cttggggaca aacatgcaac attgcagaag agcaccaagg aggttcgcag
     2461 ctcaatccgc caggtgtctg acgtacagaa gcgtgtgcaa gtggatgtca agatggccat
     2521 cctgcagatc atgaaggagc tgaataagcg gggccgtgtg ctggtcaatg atgcccagaa
     2581 ggtgactgag gggcagcagg agcgcctgga gcggcagcac tggaccatga ccaagatcca
     2641 gaagcaccag gagcacattc tgcgctttgc ctcttgggct ctggagagtg acaacaacac
     2701 agcccttttg ctttctaaga agttgatcta cttccagctg caccgggccc tcaagatgat
     2761 tgtggatccc gtggagccac atggcgagat gaagtttcag tgggacctca atgcctggac
     2821 caagagtgcc gaggcctttg gcaagattgt ggcagagcgt cctggcacta actcaacagg
     2881 ccctgcaccc atggcccctc caagagcccc agggcccctg agcaagcagg gctctggcag
     2941 cagccagccc atggaggtgc aggaaggcta tggctttggg tcaggagatg atccctactc
     3001 aagtgcagag ccccatgtgt caggtgtgaa acggtcccgc tcaggtgagg gcgaggtgag
     3061 cggccttatg cgcaaggtgc cacgagtgag ccttgaacgc ctggacctgg acctcacagc
     3121 tgacagccag ccacccgtct tcaaggtctt cccaggcagt accactgagg actacaacct
     3181 tattgttatt gaacgtggcg ctgccgctgc agctaccggc cagccaggga ctgcgcctgc
     3241 aggaacccct ggtgccccac ccctggctgg catggccatt gtcaaggagg aggagacgga
     3301 ggctgccatt ggagcccctc ctactgccac tgagggccct gagaccaaac ctgtgcttat
     3361 ggctcttgcg gagggtcctg gtgctgaggg tccccgcctg gcctcaccta gtggcagcac
     3421 cagctcaggg ctggaggtgg tggctcctga gggtacctca gccccaggtg gtggcccggg
     3481 aaccctggat gacagtgcca ccatttgccg tgtctgccag aagccaggcg atctggttat
     3541 gtgcaaccag tgtgagtttt gtttccacct ggactgtcac ctgccggccc tgcaggatgt
     3601 accaggggag gagtggagct gctcactctg ccatgtgctc cctgacctga aggaggagga
     3661 tggcagcctc agcctggatg gtgcagacag cactggcgtg gtggccaagc tctcaccagc
     3721 caaccagcgg aaatgtgagc gtgtactgct ggccctattc tgtcacgaac cctgccgccc
     3781 cctgcatcag ctggctaccg actccacctt ctccctggac cagcccggtg gcaccctgga
     3841 tctgaccctg atccgtgccc gcctccagga gaagttgtca cctccctaca gctccccaca
     3901 ggagtttgcc caggatgtgg gccgcatgtt caagcaattc aacaagttaa ctgaggacaa
     3961 ggcagacgtg cagtccatca tcggcctgca gcgcttcttc gagacgcgca tgaacgaggc
     4021 cttcggtgac accaagttct ctgctgtgct ggtggagccc ccgccgatga gcctgcctgg
     4081 tgctggcctg agttcccagg agctgtctgg tggccctggt gatggcccct gactcgagtc
     4141 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4201 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4261 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4321 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4381 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4441 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4501 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4561 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4621 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4681 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4741 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4801 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4861 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4921 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4981 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5041 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5101 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5161 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5221 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5281 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5341 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5401 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5461 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5521 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5581 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5641 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5701 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5761 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5821 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5881 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5941 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6001 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6061 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6121 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6181 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6241 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6301 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6361 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6421 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6481 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6541 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6601 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6661 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6721 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7261 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     7381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8581 gatcgggaga tctcccgatc ccctatggt
//
