LOCUS       V020815                 7358 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020815
VERSION     V020815
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"
     CDS             683..2038
                     /label="TRIM49(NM_020358)"
                     /note="TRIM49(NM_020358)"
                     /gene="TRIM49"
     CDS             2045..2110
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2134..2722
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2788..2836
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3038..3466
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3480..3809
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3876..4670
                     /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    4846..4967
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5016..5032)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5040..5056
                     /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(5064..5094)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5109..5130
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5418..6006)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6177..7037)
                     /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(7038..7142)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgaattc tggaatctta caggtctttc agggggaact
      721 catctgcccc ctgtgcatga actacttcat agacccggtc accatagact gtgggcacag
      781 cttttgcagg ccttgtttct acctcaactg gcaagacatc ccatttcttg tccagtgctc
      841 tgaatgcaca aagtcaaccg agcagataaa cctcaaaacc aacattcatt tgaagaagat
      901 ggcttctctt gccagaaaag tcagtctctg gctattcctg agctctgagg agcaaatgtg
      961 tggcactcac agggagacaa agaagatatt ctgtgaagtg gacaggagcc tgctctgttt
     1021 gctgtgctcc agctctcagg agcaccggta tcacagacac cgtcccattg agtgggctgc
     1081 tgaggaacac cgggagaagc ttttacagaa aatgcagtct ttgtgggaaa aagcttgtga
     1141 aaatcacaga aacctgaatg tggaaaccac cagaaccaga tgctggaagg attatgtgaa
     1201 tttaaggcta gaagcaatta gagctgagta tcagaagatg cctgcatttc atcatgaaga
     1261 agaaaaacat aatttggaga tgctgaaaaa gaaggggaaa gaaatttttc atcgacttca
     1321 tttaagtaaa gccaaaatgg ctcataggat ggagatttta agaggaatgt atgaggagct
     1381 gaacgaaatg tgccataaac cagatgtgga gctacttcag gcttttggag acatattaca
     1441 caggagtgag tccgtgctgc tgcacatgcc ccagcctctg aatccagagc tcagtgcagg
     1501 gcccatcact ggactgaggg acaggctcaa ccaattccga gtgcatatta ctctgcatca
     1561 tgaagaagcc aacaatgata tctttctgta tgaaattttg agaagcatgt gtattggatg
     1621 tgaccatcaa gatgtaccct atttcactgc aacacctaga agttttcttg catggggtgt
     1681 tcagactttc acctcgggca aatattactg ggaggtccat gtaggggact cctggaattg
     1741 ggcttttggt gtctgtaata tgtatcggaa agagaagaat cagaatgaga agatagatgg
     1801 aaaggcggga ctctttcttc ttgggtgtgt taagaatgac attcaatgca gtctctttac
     1861 cacctcccca cttatgctgc aatatatccc aaaacctacc agccgagtag gattattcct
     1921 ggattgtgag gctaagactg tgagctttgt tgatgttaat caaagctccc taatatacac
     1981 catccctaat tgctctttct cacctcctct caggcctatc ttttgctgta ttcacttcct
     2041 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2101 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     2161 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2221 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2281 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2341 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2401 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2461 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2521 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     2581 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2641 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2701 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     2761 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2821 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2881 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2941 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     3001 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3061 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3121 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3181 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3241 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3301 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3361 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     3421 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3481 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3721 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3781 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3841 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3901 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3961 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4021 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4081 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4141 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4201 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4261 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4321 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4381 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4441 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4501 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4561 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4621 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4681 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4741 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4801 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4981 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5581 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5821 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7141 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7201 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7261 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7321 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
