LOCUS       V046083                 7233 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046083
VERSION     V046083
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1913
                     /label="Oas1e(NM_001347450)"
                     /note="Oas1e(NM_001347450)"
                     /gene="Oas1e"
     CDS             1920..1985
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2009..2597
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2663..2711
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2913..3341
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3355..3684
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3751..4545
                     /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    4721..4842
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4891..4907)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4915..4931
                     /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(4939..4969)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4984..5005
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5293..5881)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6052..6912)
                     /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(6913..7017)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc gagggaactc ttcagaaccc
      781 caatctggag gctggataac ttcatagagg atcaactcct tcctgacacc accttcctta
      841 ctgagctcag agcagacatc gactccataa gtgctttcct gaaggagaga tgcttccaag
      901 gtgccaccca ccctatgagg gtcgccaggg ttgtgatggg aggctcctat gatgaacaca
      961 atgcactcaa gggaaggtca gaggccaaca tggtggtgtt ctttaatgat ctcaccagct
     1021 ttgaggacca gttaaagtgg cagcaagtgt tcattgaaga aattcggaaa cacctgctcc
     1081 agttgcagca agagaagcca tgtaaactca agtttgaggt gcagagctca gaggagccca
     1141 acaccaggtc tctgaccttc aagctgtgct ccccagagcg ccagcaggag gtggaatttg
     1201 atgtgcagcc agcctatgat gctctgtatg aagggggata ctgcaagtcc tttgaatcca
     1261 ttaactacaa caaagtctac gcccaactca tccatgagtg caccaccctg gagaaggagg
     1321 gcgagttctc catctgcttc accgaccttc atcagagctt cctgaggtat cgtgcgccca
     1381 agctctggaa cctcatccgt ctggtcaagc actggtatca actgtgtaag gagaagctga
     1441 ggggaccgct gcctccacag tatgccctgg agctgctcac agtctacgtc tgggaatttg
     1501 gggtccaaga cagctttgga ctccatgcag cccagtgctt ccgaacggtc ttagaactgg
     1561 tcaccaagta caaatgcctt ctaatctact ggacatggtt ttatgacttt cgacctgaga
     1621 tctctgacta cctgcacgga cagatcaaaa aagccaggcc tctgatcctg gatccggcag
     1681 acccaacaag gaacgtggct ggttcagact tacaggcatg ggacctgctg gcaaaggagg
     1741 ctcagacctg gatacattca aattttttta ggaactgtga tatgtccctt gtgaatggct
     1801 gggaagtgtc gcttccaaca gtgttctctg ggagtcaggc tgtgatggac agagaagaac
     1861 gcaagctcga cttccatctg tccacctgtt gggaggttct gtccaatagt ggcggatccg
     1921 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     1981 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2041 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2101 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2161 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2221 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2281 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2341 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2401 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2461 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2521 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2581 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     2641 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     2701 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     2761 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     2821 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     2881 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3061 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3121 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3241 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3301 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3601 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3661 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3721 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3781 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3841 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3901 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3961 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4021 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4081 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4141 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4201 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4261 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4321 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4381 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4441 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4501 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4561 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4621 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4681 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4741 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4801 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4861 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5461 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5701 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6061 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6121 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6181 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6241 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6301 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6361 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6421 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6481 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6541 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6601 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6661 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6721 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6781 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6841 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6901 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6961 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7021 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7081 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7141 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7201 ttgaccgaca attgcatgaa gaatctgctt agg
//
