LOCUS       V019704                 7100 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019704
VERSION     V019704
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             941..2632
                     /label="TRIM16(NM_006470)"
                     /note="TRIM16(NM_006470)"
                     /gene="TRIM16"
     CDS             2639..2662
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2700..2924
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2970..3398
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3412..3741
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3808..4602
                     /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    4776..4897
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4946..4962)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4970..4986
                     /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(4994..5024)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5039..5060
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5348..5933)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6104..6964)
                     /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(6965..7069)
                     /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 atggctgagt tggatctaat
      961 ggctccaggg ccactgccca gggccactgc tcagccccca gcccctctca gcccagactc
     1021 tgggtcaccc agcccagatt ctgggtcagc cagcccagtg gaagaagagg acgtgggctc
     1081 ctcggagaag cttggcaggg agacggagga acaggacagc gactctgcag agcaggggga
     1141 tcctgctggt gaggggaaag aggtcctgtg tgacttctgc cttgatgaca ccagaagagt
     1201 gaaggcagtg aagtcctgtc taacctgcat ggtgaattac tgtgaagagc acttgcagcc
     1261 gcatcaggtg aacatcaaac tgcaaagcca cctgctgacc gagccagtga aggaccacaa
     1321 ctggcgatac tgccctgccc accacagccc actgtctgcc ttctgctgcc ctgatcagca
     1381 gtgcatctgc caggactgtt gccaggagca cagtggccac accatagtct ccctggatgc
     1441 agcccgcagg gacaaggagg ctgaactcca gtgcacccag ttagacttgg agcggaaact
     1501 caagttgaat gaaaatgcca tctccaggct ccaggctaac caaaagtctg ttctggtgtc
     1561 ggtgtcagag gtcaaagcgg tggctgaaat gcagtttggg gaactccttg ctgctgtgag
     1621 gaaggcccag gccaatgtga tgctcttctt agaggagaag gagcaagctg cgctgagcca
     1681 ggccaacggt atcaaggccc acctggagta caggagtgcc gagatggaga agagcaagca
     1741 ggagctggag aggatggcgg ccatcagcaa cactgtccag ttcttggagg agtactgcaa
     1801 gtttaagaac actgaagaca tcaccttccc tagtgtttac gtagggctga aggataaact
     1861 ctcgggcatc cgcaaagtta tcacggaatc cactgtacac ttaatccagt tgctggagaa
     1921 ctataagaaa aagctccagg agttttccaa ggaagaggag tatgacatca gaactcaagt
     1981 gtctgccgtt gttcagcgca aatattggac ttccaaacct gagcccagca ccagggaaca
     2041 gttcctccaa tatgcgtatg acatcacgtt tgacccggac acagcacaca agtatctccg
     2101 gctgcaggag gagaaccgca aggtcaccaa caccacgccc tgggagcatc cctacccgga
     2161 cctccccagc aggttcctgc actggcggca ggtgctgtcc cagcagagtc tgtacctgca
     2221 caggtactat tttgaggtgg agatcttcgg ggcaggcacc tatgttggcc tgacctgcaa
     2281 aggcatcgac cggaaagggg aggagcgcaa cagttgcatt tccggaaaca acttctcctg
     2341 gagcctccaa tggaacggga aggagttcac ggcctggtac agtgacatgg agaccccact
     2401 caaagctggc cctttccgga ggctcggggt ctatatcgac ttcccgggag ggatcctttc
     2461 cttctatggc gtagagtatg ataccatgac tctggttcac aagtttgcct gcaaattttc
     2521 agaaccagtc tatgctgcct tctggctttc caagaaggaa aacgccatcc ggattgtaga
     2581 tctgggagag gaacccgaga agccagcacc gtccttggtg gggactgctc ccctcgagga
     2641 ttacaaggat gacgacgata agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     2701 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2761 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2821 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2881 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2941 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3181 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3301 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3421 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3541 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3601 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3661 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3721 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3781 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3841 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3901 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3961 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4021 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4081 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4141 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4201 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4261 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4321 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4381 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4441 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4501 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4561 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4621 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4681 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4741 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4921 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5881 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7081 gaaaagtgcc acctgacgtc
//
