LOCUS       V028844                 6818 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028844
VERSION     V028844
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"
     regulatory      935..944
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             992..2308
                     /label="DDOST(NM_005216)"
                     /note="DDOST(NM_005216)"
                     /gene="DDOST"
     CDS             2315..2380
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2418..2642
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2688..3116
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3130..3459
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3526..4320
                     /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    4494..4615
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4664..4680)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4688..4704
                     /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(4712..4742)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4757..4778
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5066..5651)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5822..6682)
                     /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(6683..6787)
                     /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 atggggtact tccggtgtgc
      961 aggtgctggg tccttcggca ggaggaggaa gatggagccc agcaccgcgg cccgggcttg
     1021 ggccctcttt tggttgctgc tgcccttgct tggcgcggtt tgcgccagcg gaccccgcac
     1081 cttagtgctg ctggacaacc tcaacgtgcg ggagactcat tcgcttttct tccggagcct
     1141 gaaggaccgg ggctttgagc tcacattcaa gaccgctgat gaccccagcc tgtctctcat
     1201 aaagtatggg gaattcctct atgacaatct catcattttc tccccttcgg tagaagattt
     1261 tggaggcaac atcaacgtgg agaccatcag tgcctttatt gacggcggag gcagtgtgct
     1321 ggtagctgcc agctccgaca ttggtgaccc tcttcgagag ctgggcagtg agtgcgggat
     1381 tgagtttgac gaggagaaaa cggctgtcat tgaccatcac aactatgaca tctcagacct
     1441 tggccagcat acgctcatcg tggctgacac tgagaacctg ctgaaggccc caaccatcgt
     1501 tgggaaatca tctctaaatc ccatcctctt tcgaggtgtt gggatggtgg ccgatcctga
     1561 taaccctttg gtgctggaca tcctgacggg ctcttccacc tcttactcct tcttcccgga
     1621 caagcctatc acccagtatc cacatgcggt ggggaagaac accctcctca ttgctgggct
     1681 ccaggccagg aacaatgccc gcgtcatctt cagcggctcc ctcgacttct tcagcgactc
     1741 cttcttcaac tcagcagtgc agaaggcggc gcccggctcc cagaggtatt cccagacagg
     1801 caactatgaa ctagctgtgg ccctctcccg ctgggtgttc aaggaggagg gtgtcctccg
     1861 tgtggggcct gtgtcccatc atcgggtggg cgagacagcc ccacccaatg cctacactgt
     1921 cactgaccta gtggagtata gcatcgtgat ccagcagctc tcaaatggca aatgggtccc
     1981 ctttgatggc gatgacattc agctggagtt tgtccgcatt gatccttttg tgaggacctt
     2041 cctgaagaag aaaggtggca aatacagtgt tcagttcaag ttgcccgacg tgtatggtgt
     2101 attccagttt aaagtggatt acaaccggct aggctacaca cacctgtact cttccactca
     2161 ggtatccgtg cggccactcc agcacacgca gtatgagcgc ttcatcccct cggcctaccc
     2221 ctactacgcc agcgccttct ccatgatgct ggggctcttc atcttcagca tcgtcttctt
     2281 gcacatgaag gagaaggaga agtccgacct cgaggactac aaagaccatg acggtgatta
     2341 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     2401 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2461 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2521 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2581 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2641 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2701 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2761 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2821 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2881 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2941 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3001 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3061 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3361 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3481 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3541 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3601 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3661 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3721 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3781 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3841 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3901 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3961 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4021 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4081 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4141 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4201 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4261 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4321 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4381 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4441 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4501 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4561 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4621 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4681 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4741 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4801 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4861 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4921 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4981 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5041 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5101 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5161 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5221 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5281 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5341 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5401 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5461 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5521 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5581 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5941 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6781 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
