LOCUS       V031996                 6720 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031996
VERSION     V031996
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..1400
                     /label="Hmgb1(NM_012963)"
                     /note="Hmgb1(NM_012963)"
                     /gene="Hmgb1"
     CDS             1407..1472
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1496..2084
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2150..2198
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2400..2828
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2842..3171
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3238..4032
                     /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    4208..4329
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4378..4394)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4402..4418
                     /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(4426..4456)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4471..4492
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4780..5368)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5539..6399)
                     /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(6400..6504)
                     /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 ccaccatggg caaaggagat cctaagaagc
      781 cgagaggcaa aatgtcctca tatgcattct ttgtgcaaac ctgccgggag gagcacaaga
      841 agaagcaccc ggatgcttct gtcaacttct cagagttctc caagaagtgc tcagagaggt
      901 ggaagaccat gtctgctaaa gaaaagggga aatttgaaga tatggcaaag gctgacaagg
      961 ctcgttatga aagagaaatg aaaacctaca tcccccccaa aggggagacc aaaaagaagt
     1021 tcaaggaccc caatgccccc aagaggcctc cttcggcctt cttcttgttc tgttctgagt
     1081 accgcccaaa aatcaaaggc gagcatcctg gcttatccat tggtgatgtt gcgaagaaac
     1141 taggagagat gtggaacaac actgctgcgg atgacaagca gccctatgaa aagaaggccg
     1201 ccaagctgaa ggagaagtat gagaaggata ttgctgccta cagagctaaa ggaaaacctg
     1261 atgcagcgaa aaagggggtg gtcaaggctg agaagagcaa gaaaaagaag gaagaggaag
     1321 acgacgagga ggatgaagag gatgaggaag aggaggaaga ggaggaagac gaagatgaag
     1381 aagaagatga tgatgatgaa ggatccgact acaaagacca tgacggtgat tataaagatc
     1441 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     1501 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     1561 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1621 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     1681 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     1741 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     1801 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1861 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     1921 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     1981 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2041 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2101 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2161 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2221 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2281 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     2341 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     2401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2461 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2521 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     2581 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2701 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2761 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2821 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2881 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3001 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3061 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3121 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3181 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3241 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3301 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3361 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3421 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3481 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3541 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3601 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3661 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3721 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3781 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3841 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3901 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3961 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4021 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4081 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4141 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4201 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4261 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4321 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4381 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4441 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4501 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4561 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4621 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4681 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4801 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     4981 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5101 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5161 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5221 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5461 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5521 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5581 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5641 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5701 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5761 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5821 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5881 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5941 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6001 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6061 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6121 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6181 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6241 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6301 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6361 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6421 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6481 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     6541 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     6601 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     6661 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
