LOCUS       V025601                 8296 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025601
VERSION     V025601
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             890..3025
                     /label="USP44(NM_032147)"
                     /note="USP44(NM_032147)"
                     /gene="USP44"
     CDS             3053..3763
                     /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"
     polyA_signal    3863..4087
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4133..4561
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4575..4904
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4971..5765
                     /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    5939..6060
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6109..6125)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6133..6149
                     /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(6157..6187)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6202..6223
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6511..7096)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7267..8127)
                     /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(8128..8232)
                     /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 cttgccacca tgctagcaat
      901 ggatacgtgc aaacatgttg ggcagctgca gcttgctcaa gaccattcca gcctcaaccc
      961 tcagaaatgg cactgtgtgg actgcaacac gaccgagtcc atttgggctt gccttagctg
     1021 ctcccatgtt gcctgtggaa gatatattga agagcatgca ctcaagcact ttcaagaaag
     1081 cagtcatcct gttgcattgg aggtgaatga gatgtacgtt ttttgttacc tttgtgatga
     1141 ttatgttctg aatgataaca caactggaga cctgaagtta ctacgacgta cattaagtgc
     1201 catcaaaagt caaaattatc actgcacaac tcgtagtggg aggtttttac ggtccatggg
     1261 tacaggtgat gattcttatt tcttacatga cggtgcccaa tctctgcttc aaagtgaaga
     1321 tcaactgtat actgctcttt ggcacaggag aaggatacta atgggtaaaa tctttcgaac
     1381 atggtttgaa caatcaccca ttggaagaaa aaagcaagaa gaaccatttc aggaaaaaat
     1441 agtagtaaaa agagaagtaa agaaaagacg gcaggaattg gagtatcaag ttaaagcaga
     1501 attggaaagt atgcctccaa gaaagagttt acgtttacaa gggctcgctc agtcgaccat
     1561 aatagaaata gtttctgttc aggtgccagc acaaacgcca gcatcaccag caaaagataa
     1621 agtactctct acctcagaaa atgaaatatc tcaaaaagtc agtgactcct cagttaaacg
     1681 aaggccaata gtaactcctg gtgtaacagg attgagaaat ttgggaaata cttgctatat
     1741 gaattctgtt cttcaggtgt tgagtcattt acttattttt cgacaatgtt ttttaaagct
     1801 tgatctgaac caatggctgg ctatgactgc tagcgagaag acaagatctt gtaagcatcc
     1861 accagtcaca gatacagtag tatatcaaat gaatgaatgt caggaaaaag atacaggttt
     1921 tgtttgctcc agacaatcaa gtctgtcatc aggactaagt ggtggagcat caaaaggtag
     1981 aaagatggaa cttattcagc caaaggagcc aacttcacag tacatttctc tttgtcatga
     2041 attgcatact ttgttccaag tcatgtggtc tggaaagtgg gcgttggtct caccatttgc
     2101 tatgctacac tcagtgtgga gactcattcc tgcctttcgt ggttacgccc aacaagacgc
     2161 tcaggaattt ctttgtgaac ttttagataa aatacaacgt gaattagaga caactggtac
     2221 cagtttacca gctcttatcc ccacttctca aaggaaactc atcaaacaag ttctgaatgt
     2281 tgtaaataac atttttcatg gacaacttct tagtcaggtt acatgtcttg catgtgacaa
     2341 caaatcaaat accatagaac ctttctggga cttgtcattg gagtttccag aaaggtatca
     2401 atgcagtgga aaagatattg cttcccagcc atgtctggtt actgaaatgt tggccaaatt
     2461 tacagaaact gaagctttag aaggaaaaat ctacgtatgt gaccagtgta actcaaagcg
     2521 tagaaggttt tcctccaaac cagttgtact cacagaagcc cagaaacaac ttatgatatg
     2581 ccacctacct caggttctca gactgcacct caaacgattc aggtggtcag gacgtaataa
     2641 ccgagagaag attggtgttc atgttggctt tgaggaaatc ttaaacatgg agccctattg
     2701 ctgcagggag accctgaaat ccctcagacc agaatgcttt atctatgact tgtccgcggt
     2761 ggtgatgcac catgggaaag gatttggctc agggcactac actgcctact gctataattc
     2821 tgaaggaggg ttctgggtac actgcaatga ttccaaacta agcatgtgca ctatggatga
     2881 agtatgcaag gctcaagctt atatcttgtt ttatacccaa cgagttactg agaatggaca
     2941 ttctaaactt ttgcctccag agctcctgtt ggggagccaa catcccaatg aagacgctga
     3001 tacctcgtct aatgaaatcc ttagcggagg tggcggaagt ggtaccgcca ccatggtgag
     3061 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     3121 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3181 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     3241 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     3301 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3361 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     3421 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     3481 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     3541 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     3601 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     3661 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     3721 gggccgccac tccaccggcg gcatggacga gctgtacaag tagggatcca ctagtccagt
     3781 gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     3841 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3901 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3961 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4021 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4081 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4141 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4201 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4261 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4321 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4381 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4441 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4501 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4561 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4621 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4741 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4801 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4861 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4921 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4981 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5041 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5101 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5161 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5221 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5281 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5341 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5401 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5461 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5521 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5581 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5641 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5701 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5761 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5821 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5881 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6061 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7021 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     7081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8281 cccgatcccc tatggt
//
