LOCUS       V025691                 7179 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025691
VERSION     V025691
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..1859
                     /label="Usp18(NM_011909)"
                     /note="Usp18(NM_011909)"
                     /gene="Usp18"
     CDS             1866..1931
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1955..2543
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2609..2657
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2859..3287
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3301..3630
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3697..4491
                     /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    4667..4788
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4837..4853)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4861..4877
                     /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(4885..4915)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4930..4951
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5239..5827)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5998..6858)
                     /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(6859..6963)
                     /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 ccaccatggg caaggggttt gggctcctga
      781 ggaaaccctg ccagtcagtt gtggctgagc cgcagcagta ctcagcgctg gaggaagaga
      841 ggaccatgaa gaggaagaga gtgctgtcta gagacctctg cagtgcctgg gacagccctc
      901 atggtctggt tggtttacac aacatcggac agacgtgttg ccttaactcc ttgcttcaag
      961 tgttcatgat gaacatggac ttcagaatga tattgaagag gataacagtg cctcggagtg
     1021 cagaagagag gaagagaagt gtccccttcc agctgctttt gctgctggag aagatgcagg
     1081 acagtcgaca gaaagccgtg ctgcccacgg agctcgttca gtgcctgcag aaatacaacg
     1141 tgccgttgtt tgtccagcac gatgctgctc aactctacct tacaatctgg aacctgacta
     1201 aggaccagat cacggacaca gacttgacag agcggctgca gggtctgttc accatctgga
     1261 cgcaggagtc cctgatttgc gtgggctgta cagcagagag cagcaggagg agcaaactgc
     1321 tcacactctc tcttcctctt tttgataagg acgcaaagcc tctgaaaacc ctggaggatg
     1381 ccttgagatg tttcgtccag cccaaagagt tagcaagctc cgacatgtgc tgcgagagct
     1441 gtggggagaa gacgccttgg aaacaggttc tgaagctgac tcatctgccc caaaccttga
     1501 ccattcacct catgagattc tctgccagga actcgaggac ggaaaagatc tgtcactcag
     1561 ttaatttccc tcagagcttg gatttcagtc aggttctgcc aaccgaggaa gacctcggtg
     1621 ataccaagga acagtctgaa atacactatg aactctttgc cgtgattgcc cacgttggga
     1681 tggctgactt tggtcattac tgtgcctaca tccggaatcc cgtggatgga aagtggttct
     1741 gcttcaatga ctcacatgtt tgttgggtca cctggaagga tgtccagtgt acctatggga
     1801 accacagata ccgctggcgg gaaacggcct atctcttggt ttacacgaag actggatccg
     1861 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     1921 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     1981 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2041 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2101 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2161 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2221 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2281 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2341 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2401 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2461 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2521 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     2581 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2641 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     2701 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     2761 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     2821 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2881 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2941 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3001 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3061 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3121 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3181 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3241 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3301 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3421 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3541 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3601 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3661 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3721 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3781 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3841 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3901 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3961 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4021 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4081 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4141 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4201 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4261 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4321 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4381 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4441 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4501 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4561 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4621 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4681 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4741 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4801 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5041 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5101 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5161 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5221 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5281 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5341 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5401 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5461 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5521 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5581 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5641 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5701 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5761 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5821 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5881 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5941 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6001 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6061 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6121 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6961 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7021 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7081 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7141 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
