LOCUS       V043915                 6679 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043915
VERSION     V043915
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2169
                     /label="Tbc1d20(NM_024196)"
                     /note="Tbc1d20(NM_024196)"
                     /gene="Tbc1d20"
     CDS             2176..2241
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2279..2503
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2549..2977
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2991..3320
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3387..4181
                     /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    4355..4476
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4525..4541)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4549..4565
                     /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(4573..4603)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4618..4639
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4927..5512)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5683..6543)
                     /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(6544..6648)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggccc tccggccctc aaagggcgac ggctccgccg gccgctggga ccgcggcgca
     1021 gggaaggcag actttaatgc caaaaggaaa aagaaggtgg cagagataca ccaggctctg
     1081 aacagtgatc ccatcgactt ggctgccctc cgacgaatgg ctatcagtga gggagggctc
     1141 ctgactgatg agatcaggtg ccaagtatgg cctaagctcc tcaacgtgaa caccagtgag
     1201 ccaccgcctg tgtcaaggaa ggaccttcga gacatgagca aggactacca gcaagtgctg
     1261 ctggacgtca ggcgatctct tcggcgcttc ccccctggca tgccggacga gcagagagaa
     1321 gggctccagg aagagctaat cgatatcatc ctcctcgtcc tggatcgcaa ccctcagctc
     1381 cactactacc agggctacca tgacatcgtg gtcacatttc tgctggtggt gggggagagg
     1441 ctggcaacat ccctggtaga aaaattgtct acccaccacc tcagggattt catggatccc
     1501 acaatggaca acaccaagca catcctaaat tatctgatgc ccatcattga ccaagtgagc
     1561 ccagagctcc acgacttcat gcagagtgct gaggtgggga ccatctttgc cctcagctgg
     1621 cttatcacct ggtttgggca cgtcctgatg gacttcaggc acgttgtgag gttatacgac
     1681 ttcttcctgg cctgccaccc acttatgccc atttactttg cagctgtgat cgtgctgtac
     1741 cgggagcagg aagtcctgga ctgtgactgt gacatggcct ctgtccacca cctgttgtcc
     1801 cagatccctc aggacctgcc ctatgagacg ctcatcagca gagcaggaga cctctttgtt
     1861 cagtttcccc cttctgaact tgcaagggag gcagctgctc agcaggaggc tgagagaacg
     1921 gcagcctcta ctttcaaaga ctttgagctg gcatcaactc agcagaggcc agacatggtg
     1981 ctgcggcagc ggtttcgggg acttctgagg cctgaggctc gaacaaaaga tgtcctgacc
     2041 aaaccaagga ccaaccgctt tgtgaaactg gcagtgatgg ggctgacggt ggccctcgga
     2101 gcagcagcac tagcagtggt gaagagcgcc ctggagtggg cccctaagtt ccagctgcag
     2161 ctgttccccc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     2221 tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc agcctcgact
     2281 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2341 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2401 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2461 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2521 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2941 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3361 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3421 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3481 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3541 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3601 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3661 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3721 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3781 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3841 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3901 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3961 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4021 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4081 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4141 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4201 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4261 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4321 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4381 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4441 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4501 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4681 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5461 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6661 aaaagtgcca cctgacgtc
//
