LOCUS       V023520                 7920 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023520
VERSION     V023520
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..2600
                     /label="Trmt2a(NM_001011895)"
                     /note="Trmt2a(NM_001011895)"
                     /gene="Trmt2a"
     CDS             2607..2672
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2696..3284
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3350..3398
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3600..4028
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4042..4371
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4438..5232
                     /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    5408..5529
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5578..5594)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5602..5618
                     /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(5626..5656)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5671..5692
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5980..6568)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6739..7599)
                     /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(7600..7704)
                     /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 ctcggatccg ccaccatgag tgagaagctg gacacggaag
      781 ttccggagca catggaggac tgcggccagg atgtcagtgc agtgggcagc tctgcagcct
      841 cagtatgtca agatgagaaa gaggaccctg ggccagctgc agggcctgga actcagcctg
      901 ggctctacag ctacataagg gatgacttgt tcacctcaga gatctttaaa ctggagctcc
      961 aaaatgtacc tcgccacgcc agcttcagtg acgtccggcg ttttttaggc cgttttggct
     1021 tacaatccca caaaatcaaa ctctttggac agccaccatg tgcctttgtg acatttcgca
     1081 gtgctgctga acgagacaag gccttgcgag tgctgcatgg tgttctctgg aagggtcgtc
     1141 cactcagtgt acgactggcc cgacccaagg ctgaccccat ggctaggaag aggcggcaag
     1201 aaggcgataa tgaaccaaca gcaacacaga ttgccgatgt ggtgacccct ctttggacag
     1261 tgccctacac tgagcagctg gagcagaagc gactggaatg tgagcgggtg ctacagaaac
     1321 tggccaagga aattgggaac actaaccatg ccctgctacc ctggctgctc ttgcagagac
     1381 aacagcacaa taaagcttgt tgcccattgg agggggtcaa gccatcaccc cagcagactg
     1441 agtaccgtaa caagtgtgaa tttctggtcg gagttggggt agatggagag gacaacactg
     1501 ttggctgccg gcttggcaag tacaagggcg ggacgtgtgc tgtggcgcct ccctttgaca
     1561 ctgtgcacat tccacaggcc actaagcagc tggtgaaggc cttccaggag ttcatccgtt
     1621 ccacaccata ttcggcatat gaccctgaga catacacagg ccactggaag caactgactg
     1681 tccgcaccag ccgccgaggt caagccatgg ccattgccca cttccatccc cagaaactga
     1741 gctctgagga ggtggcagga ctaaaggcct ccctggtgtg ccacttcatg gaggggccag
     1801 gcaaggccag tggggtgaca tccctctact ttgtggagga gggacagaga aagactccca
     1861 gccaggaagg cctgcccctg gagcatatgg ccggtgacca gtgcattcag gaagacctgc
     1921 tagggctcac tttccgcatc tcccctcatg cattcttcca ggtaaacaca cctgcagccg
     1981 aggtgctcta cacagtcatc caggaatggg cccagctgga tgggggcagc acagtgttgg
     2041 acgtttgctg tggcactggt accatagggc tagccctggc tccgaaggtg aagaaagtaa
     2101 ttgggattga actgtgccag gaggctgtgg aggatgcccg tatgaacgcc ctcaccaatg
     2161 agctgagcaa cgttgagttc cactgcggca gggctgagga tctggtgcca ggcttggtga
     2221 gcaaactgtc ttcccagcaa cttgtagctg ttctagaccc accacgggct ggcctacatt
     2281 ctaaggtgat tctggccatg aggaaagctg agaacatcaa gcggctcctg tatgtttcct
     2341 gcaaccccag ggcagccatg ggcaactttg tggatctctg cagagcccca tctaaccgag
     2401 tgaaaggtac cccattccat ccagtcaagg ctgtggcagt ggacctgttc ccacagaccc
     2461 cacactgtga aatgcttatc ctgtttgaga ggatgcaaca acaccctaat ggcatagggg
     2521 ccctggaaca tcaagacctt caaaccccaa gaaatcttcc cgacctcact gcacaggaaa
     2581 cagagacttc cctctcaccc ctcgaggact acaaagacca tgacggtgat tataaagatc
     2641 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2701 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2761 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2821 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2881 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2941 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3001 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3061 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3121 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3181 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3241 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3301 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3361 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3421 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3481 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3541 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3961 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4201 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4261 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4321 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4381 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4441 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4501 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4561 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4621 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4681 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4741 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4801 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4861 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4921 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4981 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5041 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5101 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5161 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5221 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5281 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5341 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5401 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5461 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5521 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5581 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5641 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5701 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5761 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5821 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5881 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5941 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6001 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6061 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6121 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6181 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6241 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6301 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6361 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6421 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6481 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6541 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6601 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6661 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6721 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6781 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6841 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6901 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6961 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7021 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7081 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7141 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7201 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7261 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7321 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7381 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7441 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7501 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7561 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7621 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7681 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7741 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7801 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7861 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
