LOCUS       V034623                 7258 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034623
VERSION     V034623
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..2748
                     /label="GBP3(NM_018284)"
                     /note="GBP3(NM_018284)"
                     /gene="GBP3"
     CDS             2755..2820
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2858..3082
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3128..3556
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3570..3899
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3966..4760
                     /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    4934..5055
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5104..5120)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5128..5144
                     /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(5152..5182)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5197..5218
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5506..6091)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6262..7122)
                     /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(7123..7227)
                     /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 accatggctc cagagatcca catgacaggc ccaatgtgcc tcattgagaa cactaatggg
     1021 gaactggtgg cgaatccaga agctctgaaa atcctgtctg ccattacaca gcctgtggtg
     1081 gtggtggcaa ttgtgggcct ctaccgcaca ggaaaatcct acctgatgaa caagctagct
     1141 gggaagaata agggcttctc tctgggctcc acagtgaaat ctcacaccaa aggaatctgg
     1201 atgtggtgtg tgcctcaccc caaaaagcca gaacacacct tagtcctgct tgacactgag
     1261 ggcctgggag atgtaaagaa gggtgacaac cagaatgact cctggatctt caccctggcc
     1321 gtcctcctga gcagcactct cgtgtacaat agcatgggaa ccatcaacca gcaggctatg
     1381 gaccaactgt actatgtgac agagctgaca catcgaatcc gatcaaaatc ctcacctgat
     1441 gagaatgaga atgaggattc agctgacttt gtgagcttct tcccagattt tgtgtggaca
     1501 ctgagagatt tctccctgga cttggaagca gatggacaac ccctcacacc agatgagtac
     1561 ctggagtatt ccctgaagct aacgcaaggt accagtcaaa aagataaaaa ttttaatctg
     1621 ccccgactct gtatccggaa gttcttccca aagaaaaaat gttttgtctt cgatctgccc
     1681 attcaccgca ggaagcttgc ccagcttgag aaactacaag atgaagagct ggaccctgaa
     1741 tttgtgcaac aagtagcaga cttctgttcc tacatcttta gcaattccaa aactaaaact
     1801 ctttcaggag gcatcaaggt caatgggcct cgtctagaga gcctagtgct gacctatatc
     1861 aatgctatca gcagagggga tctgccctgc atggagaacg cagtcctggc cttggcccag
     1921 atagagaact cagccgcagt gcaaaaggct attgcccact atgaccagca gatgggccag
     1981 aaggtgcagc tgcccgcaga aaccctccag gagctgctgg acctgcacag ggttagtgag
     2041 agggaggcca ctgaagtcta tatgaagaac tctttcaagg atgtggacca tctgtttcaa
     2101 aagaaattag cggcccagct agacaaaaag cgggatgact tttgtaaaca gaatcaagaa
     2161 gcatcatcag atcgttgctc agctttactt caggtcattt tcagtcctct agaagaagaa
     2221 gtgaaggcgg gaatttattc gaaaccaggg ggctattgtc tctttattca gaagctacaa
     2281 gacctggaga aaaagtacta tgaggaacca aggaagggga tacaggctga agagattctg
     2341 cagacatact tgaaatccaa ggagtctgtg accgatgcaa ttctacagac agaccagatt
     2401 ctcacagaaa aggaaaagga gattgaagtg gaatgtgtaa aagctgaatc tgcacaggct
     2461 tcagcaaaaa tggtggagga aatgcaaata aagtatcagc agatgatgga agagaaagag
     2521 aagagttatc aagaacatgt gaaacaattg actgagaaga tggagaggga gagggcccag
     2581 ttgctggaag agcaagagaa gaccctcact agtaaacttc aggaacaggc ccgagtacta
     2641 aaggagagat gccaaggtga aagtacccaa cttcaaaatg agatacaaaa gctacagaag
     2701 accctgaaaa aaaaaaccaa gagatatatg tcgcataagc taaagatcct cgaggactac
     2761 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     2821 tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2881 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2941 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3001 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3061 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3121 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3181 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3241 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3301 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3361 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3421 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3481 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3541 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3661 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3721 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3901 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3961 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4021 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4081 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4141 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4201 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4261 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4321 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4381 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4441 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4501 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4561 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4621 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4681 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4741 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4801 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4861 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4921 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4981 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5041 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5941 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6061 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6121 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6181 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6241 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6301 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6361 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6421 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6481 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6541 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6601 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6661 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6721 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6781 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6841 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6901 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6961 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7021 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7081 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7141 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7201 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
