LOCUS       V025805                 9249 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025805
VERSION     V025805
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..3929
                     /label="UBA1(NM_003334)"
                     /note="UBA1(NM_003334)"
                     /gene="UBA1"
     CDS             3936..4001
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4025..4613
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4679..4727
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4929..5357
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5371..5700
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5767..6561
                     /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    6737..6858
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6907..6923)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6931..6947
                     /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(6955..6985)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7000..7021
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7309..7897)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8068..8928)
                     /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(8929..9033)
                     /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 ccaccatgtc cagctcgccg ctgtccaaga
      781 aacgtcgcgt gtccgggcct gatccaaagc cgggttctaa ctgctcccct gcccagtccg
      841 tgttgtccga agtgccctcg gtgccaacca acggaatggc caagaacggc agtgaagcag
      901 acatagacga gggcctttac tcccggcagc tgtatgtgtt gggccatgag gcaatgaagc
      961 ggctccagac atccagtgtc ctggtatcag gcctgcgggg cctgggcgtg gagatcgcta
     1021 agaacatcat ccttggtggg gtcaaggctg ttaccctaca tgaccagggc actgcccagt
     1081 gggctgatct ttcctcccag ttctacctgc gggaggagga catcggtaaa aaccgggccg
     1141 aggtatcaca gccccgcctc gctgagctca acagctatgt gcctgtcact gcctacactg
     1201 gacccctcgt tgaggacttc cttagtggtt tccaggtggt ggtgctcacc aacacccccc
     1261 tggaggacca gctgcgagtg ggtgagttct gtcacaaccg tggcatcaag ctggtggtgg
     1321 cagacacgcg gggcctgttt gggcagctct tctgtgactt tggagaggaa atgatcctca
     1381 cagattccaa tggggagcag ccactcagtg ctatggtttc tatggttacc aaggacaacc
     1441 ccggtgtggt tacctgcctg gatgaggccc gacacgggtt tgagagcggg gactttgtct
     1501 ccttttcaga agtacagggc atggttgaac tcaacggaaa tcagcccatg gagatcaaag
     1561 tcctgggtcc ttataccttt agcatctgtg acacctccaa cttctccgac tacatccgtg
     1621 gaggcatcgt cagtcaggtc aaagtaccta agaagattag ctttaaatcc ttggtggcct
     1681 cactggcaga acctgacttt gtggtgacgg acttcgccaa gttttctcgc cctgcccagc
     1741 tgcacattgg cttccaggcc ctgcaccagt tctgtgctca gcatggccgg ccacctcggc
     1801 cccgcaatga ggaggatgca gcagaactgg tagccttagc acaggctgtg aatgctcgag
     1861 ccctgccagc agtgcagcaa aataacctgg acgaggacct catccggaag ctggcatatg
     1921 tggctgctgg ggatctggca cccataaacg ccttcattgg gggcctggct gcccaggaag
     1981 tcatgaaggc ctgctccggg aagttcatgc ccatcatgca gtggctatac tttgatgccc
     2041 ttgagtgtct ccctgaggac aaagaggtcc tcacagagga caagtgcctc cagcgccaga
     2101 accgttatga cgggcaagtg gctgtgtttg gctcagacct gcaagagaag ctgggcaagc
     2161 agaagtattt cctggtgggt gcgggggcca ttggctgtga gctgctcaag aactttgcca
     2221 tgattgggct gggctgcggg gagggtggag aaatcatcgt tacagacatg gacaccattg
     2281 agaagtcaaa tctgaatcga cagtttcttt tccggccctg ggatgtcacg aagttaaagt
     2341 ctgacacggc tgctgcagct gtgcgccaaa tgaatccaca tatccgggtg acaagccacc
     2401 agaaccgtgt gggtcctgac acggagcgca tctatgatga cgattttttc caaaacctag
     2461 atggcgtggc caatgccctg gacaacgtgg atgcccgcat gtacatggac cgccgctgtg
     2521 tctactaccg gaagccactg ctggagtcag gcacactggg caccaaaggc aatgtgcagg
     2581 tggtgatccc cttcctgaca gagtcgtaca gttccagcca ggacccacct gagaagtcca
     2641 tccccatctg taccctgaag aacttcccta atgccatcga gcacaccctg cagtgggctc
     2701 gggatgagtt tgaaggcctc ttcaagcagc cagcagaaaa tgtcaaccag tacctcacag
     2761 accccaagtt tgtggagcga acactgcggc tggcaggcac tcagcccttg gaggtgctgg
     2821 aggctgtgca gcgcagcctg gtgctgcagc gaccacagac ctgggctgac tgcgtgacct
     2881 gggcctgcca ccactggcac acccagtact cgaacaacat ccggcagctg ctgcacaact
     2941 tccctcctga ccagctcaca agctcaggag cgccgttctg gtctgggccc aaacgctgtc
     3001 cacacccgct cacctttgat gtcaacaatc ccctgcatct ggactatgtg atggctgctg
     3061 ccaacctgtt tgcccagacc tacgggctga caggctctca ggaccgagct gctgtggcca
     3121 cattcctgca gtctgtgcag gtccccgaat tcacccccaa gtctggcgtc aagatccatg
     3181 tttctgacca ggagctgcag agcgccaatg cctctgttga tgacagtcgt ctagaggagc
     3241 tcaaagccac tctgcccagc ccagacaagc tccctggatt caagatgtac cccattgact
     3301 ttgagaagga tgatgacagc aactttcata tggatttcat cgtggctgca tccaacctcc
     3361 gggcagaaaa ctatgacatt ccttctgcag accggcacaa gagcaagctg attgcaggga
     3421 agatcatccc agccattgcc acgaccacag cagccgtggt tggccttgtg tgtctggagc
     3481 tgtacaaggt tgtgcagggg caccgacagc ttgactccta caagaatggt ttcctcaact
     3541 tggccctgcc tttctttggt ttctctgaac cccttgccgc accacgtcac cagtactata
     3601 accaagagtg gacattgtgg gatcgctttg aggtacaagg gctgcagcct aatggtgagg
     3661 agatgaccct caaacagttc ctcgactatt ttaagacaga gcacaaatta gagatcacca
     3721 tgctgtccca gggcgtgtcc atgctctatt ccttcttcat gccagctgcc aagctcaagg
     3781 aacggttgga tcagccgatg acagagattg tgagccgtgt gtcgaagcga aagctgggcc
     3841 gccacgtgcg ggcgctggtg cttgagctgt gctgtaacga cgagagcggc gaggatgtcg
     3901 aggttcccta tgtccgatac accatccgcg gatccgacta caaagaccat gacggtgatt
     3961 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     4021 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4081 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4141 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4201 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4261 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4321 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4381 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4441 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4501 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4561 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4621 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4681 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4741 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4801 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4861 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4921 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4981 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5041 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     5101 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5161 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5221 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5281 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     5341 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5401 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5521 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5581 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5641 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5701 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5761 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5821 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5881 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5941 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6001 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6061 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6121 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6181 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6241 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6301 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6361 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6421 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6481 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6541 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6601 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6661 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6721 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6781 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6841 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7501 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7741 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9061 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     9121 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9181 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9241 ctgcttagg
//
