LOCUS       V040511                 8434 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040511
VERSION     V040511
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"
     CDS             706..3093
                     /label="VPS35(NM_018206)"
                     /note="VPS35(NM_018206)"
                     /gene="VPS35"
     CDS             3100..3126
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3130..3156
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3160..3186
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3210..3798
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3864..3912
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4114..4542
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4556..4885
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4952..5746
                     /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    5922..6043
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6092..6108)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6116..6132
                     /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(6140..6170)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6185..6206
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6494..7082)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7253..8113)
                     /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(8114..8218)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgcc tacaacacag
      721 cagtcccctc aggatgagca ggaaaagctc ttggatgaag ccatacaggc tgtgaaggtc
      781 cagtcattcc aaatgaagag atgcctggac aaaaacaagc ttatggatgc tctaaaacat
      841 gcttctaata tgcttggtga actccggact tctatgttat caccaaagag ttactatgaa
      901 ctttatatgg ccatttctga tgaactgcac tacttggagg tctacctgac agatgagttt
      961 gctaaaggaa ggaaagtggc agatctctac gaacttgtac agtatgctgg aaacattatc
     1021 ccaaggcttt accttttgat cacagttgga gttgtatatg tcaagtcatt tcctcagtcc
     1081 aggaaggata ttttgaaaga tttggtagaa atgtgccgtg gtgtgcaaca tcccttgagg
     1141 ggtctgtttc ttcgaaatta ccttcttcag tgtaccagaa atatcttacc tgatgaagga
     1201 gagccaacag atgaagaaac aactggtgac atcagtgatt ccatggattt tgtactgctc
     1261 aactttgcag aaatgaacaa gctctgggtg cgaatgcagc atcagggaca tagccgagat
     1321 agagaaaaaa gagaacgaga aagacaagaa ctgagaattt tagtgggaac aaatttggtg
     1381 cgcctcagtc agttggaagg tgtaaatgtg gaacgttaca aacagattgt tttgactggc
     1441 atattggagc aagttgtaaa ctgtagggat gctttggctc aagaatatct catggagtgt
     1501 attattcagg ttttccctga tgaatttcac ctccagactt tgaatccttt tcttcgggcc
     1561 tgtgctgagt tacaccagaa tgtaaatgtg aagaacataa tcattgcttt aattgataga
     1621 ttagctttat ttgctcaccg tgaagatgga cctggaatcc cagcggatat taaacttttt
     1681 gatatatttt cacagcaggt ggctacagtg atacagtcta gacaagacat gccttcagag
     1741 gatgttgtat ctttacaagt ctctctgatt aatcttgcca tgaaatgtta ccctgatcgt
     1801 gtggactatg ttgataaagt tctagaaaca acagtggaga tattcaataa gctcaacctt
     1861 gaacatattg ctaccagtag tgcagtttca aaggaactca ccagactttt gaaaatacca
     1921 gttgacactt acaacaatat tttaacagtc ttgaaattaa aacattttca cccactcttt
     1981 gagtactttg actacgagtc cagaaagagc atgagttgtt atgtgcttag taatgttctg
     2041 gattataaca cagaaattgt ctctcaagac caggtggatt ccataatgaa tttggtatcc
     2101 acgttgattc aagatcagcc agatcaacct gtagaagacc ctgatccaga agattttgct
     2161 gatgagcaga gccttgtggg ccgcttcatt catctgctgc gctctgagga ccctgaccag
     2221 cagtacttga ttttgaacac agcacgaaaa cattttggag ctggtggaaa tcagcggatt
     2281 cgcttcacac tgccaccttt ggtatttgca gcttaccagc tggcttttcg atataaagag
     2341 aattctaaag tggatgacaa atgggaaaag aaatgccaga agattttttc atttgcccac
     2401 cagactatca gtgctttgat caaagcagag ctggcagaat tgcccttaag actttttctt
     2461 caaggagcac tagctgctgg ggaaattggt tttgaaaatc atgagacagt cgcatatgaa
     2521 ttcatgtccc aggcattttc tctgtatgaa gatgaaatca gcgattccaa agcacagcta
     2581 gctgccatca ccttgatcat tggcactttt gaaaggatga agtgcttcag tgaagagaat
     2641 cacgaacctc tgaggactca gtgtgccctt gctgcatcca aacttctaaa gaaacctgat
     2701 cagggccgag ctgtgagcac ctgtgcacat ctcttctggt ctggcagaaa cacggacaaa
     2761 aatggggagg agcttcacgg aggcaagagg gtaatggagt gcctaaaaaa agctctaaaa
     2821 atagcaaatc agtgcatgga cccctctcta caagtgcagc tttttataga aattctgaac
     2881 agatatatct atttttatga aaaggaaaat gatgcggtaa caattcaggt tttaaaccag
     2941 cttatccaaa agattcgaga agacctcccg aatcttgaat ccagtgaaga aacagagcag
     3001 attaacaaac attttcataa cacactggag catttgcgct tgcggcggga atcaccagaa
     3061 tccgaggggc caatttatga aggtctcatc cttctcgagt acccatacga cgtacctgat
     3121 tatgcaactt atccttatga tgtacctgat tatgctagct atccttatga tgtcccagac
     3181 tacgcatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     3241 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3301 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3361 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3421 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3481 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3541 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3601 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3661 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3721 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3781 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     3841 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     3901 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3961 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4021 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     4081 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4141 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4201 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4261 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4321 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4381 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4441 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4501 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4561 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4621 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4681 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4741 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4801 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4861 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4921 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4981 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5041 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5101 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5161 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5221 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5281 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5341 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5401 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5461 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5521 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5581 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5641 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5701 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5761 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5821 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5881 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5941 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6001 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6061 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6661 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6901 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8221 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8281 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     8341 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     8401 cttgaccgac aattgcatga agaatctgct tagg
//
