LOCUS       V028132                 7457 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028132
VERSION     V028132
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             1904..2947
                     /label="Rho(NM_145383)"
                     /note="Rho(NM_145383)"
                     /gene="Rho"
     CDS             2954..3019
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3057..3281
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3327..3755
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3769..4098
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4165..4959
                     /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    5133..5254
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5303..5319)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5327..5343
                     /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(5351..5381)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5396..5417
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5705..6290)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6461..7321)
                     /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(7322..7426)
                     /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 atccgccacc atgataactt ttctgcccat
      961 cattttttcc attctaatag tggttacatt tgtgattgga aattttgcta atggcttcat
     1021 agcattggta aattccattg agtggttcaa gagacaaaag atctcttttg ctgaccaaat
     1081 tctcactgct ctggcagtct ccagagttgg tttactctgg gtattagtat taaattggta
     1141 tgcaactgag ttgaatccag cttttaacag tatagaagta agaattactg cttacaatgt
     1201 ctgggcagta atcaaccatt tcagcaactg gcttgctact agcctcagca tattttattt
     1261 gctcaagatt gccaatttct ccaaccttat ttttcttcac ttaaagagga gagttaagag
     1321 tgttgttctg gtgatactat tggggccttt gctatttttg gtttgtcatc tttttgtgat
     1381 aaacatgaat cagattatat ggacaaaaga atatgaagga aacatgactt ggaagatcaa
     1441 actgaggagt gcaatgtacc tttcaaatac aacggtaacc atcctagcaa acttagttcc
     1501 cttcactctg accctgatat cttttctgct gttaatctgt tctctgtgta aacatctcaa
     1561 aaagatgcag ctccatggca aaggatctca agatcccagc atgaaggtcc acataaaagc
     1621 tttgcaaact gtgacctcct tcctcttgtt atgtgccatt tactttctgt ccataatcat
     1681 gtcagtttgg agttttgaga gtctggaaaa caaacctgtc ttcatgttct gcgaagctat
     1741 tgcattcagc tatccttcaa cccacccatt catcctgatt tggggaaaca agaagctaaa
     1801 gcagactttt ctttcagttt tgtggcatgt gaggtactgg gtgaaaggag agaagccttc
     1861 atcttcaggt ggaggcggat caggaggcgg tggatctgaa ttcatgaacg gcacagaggg
     1921 ccccaatttt tatgtgccct tctccaacgt cacaggcgtg gtgcggagcc ccttcgagca
     1981 gccgcagtac tacctggcgg aaccatggca gttctccatg ctggcagcgt acatgttcct
     2041 gctcatcgtg ctgggcttcc ccatcaactt cctcacgctc tacgtcaccg tacagcacaa
     2101 gaagctgcgc acacccctca actacatcct gctcaacttg gccgtggctg acctcttcat
     2161 ggtcttcgga ggattcacca ccaccctcta cacatcactc catggctact tcgtctttgg
     2221 gcccacaggc tgtaatctcg agggcttctt tgccacactt ggaggtgaaa tcgccctgtg
     2281 gtccctggtg gtcctggcca ttgagcgcta cgtggtggtc tgcaagccga tgagcaactt
     2341 ccgcttcggg gagaatcacg ctatcatggg tgtggtcttc acctggatca tggcgttggc
     2401 ctgtgctgct cccccactcg ttggctggtc caggtacatc cctgagggca tgcaatgttc
     2461 atgcgggatt gactactaca cactcaagcc tgaggtcaac aacgaatcct ttgtcatcta
     2521 catgttcgtg gtccacttca ccattcctat gatcgtcatc ttcttctgct atgggcagct
     2581 ggtcttcaca gtcaaggagg cggctgccca gcagcaggag tcagccacca ctcagaaggc
     2641 agagaaggaa gtcacccgca tggttatcat catggtcatc ttcttcctga tctgctggct
     2701 tccctacgcc agtgtggcct tctacatctt cacccaccag ggctccaact tcggccccat
     2761 cttcatgact ctgccagctt tctttgctaa gagctcttcc atctataacc cggtcatcta
     2821 catcatgttg aacaagcagt tccggaactg tatgctcacc acgctgtgct gcggcaagaa
     2881 tccactggga gatgacgacg cctctgccac cgcttccaag acggagacca gccaggtggc
     2941 tccagccctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3001 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     3061 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3121 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3181 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3241 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3301 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3361 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3421 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3481 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3541 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3601 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3661 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3721 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3781 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3841 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3901 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3961 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4021 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4081 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4141 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4201 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4261 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4321 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4381 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4441 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4501 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4561 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4621 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4681 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4741 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4801 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4861 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4921 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4981 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5041 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5101 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5161 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5221 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5281 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5341 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5401 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5461 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5521 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5581 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5641 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5701 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5761 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5821 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5881 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5941 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6001 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6061 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6121 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6181 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7441 aagtgccacc tgacgtc
//
