LOCUS       V025907                 6921 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025907
VERSION     V025907
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..1601
                     /label="HMGN5(NM_030763)"
                     /note="HMGN5(NM_030763)"
                     /gene="HMGN5"
     CDS             1608..1673
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1697..2285
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2351..2399
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2601..3029
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3043..3372
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3439..4233
                     /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    4409..4530
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4579..4595)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4603..4619
                     /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(4627..4657)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4672..4693
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4981..5569)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5740..6600)
                     /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(6601..6705)
                     /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 ccaccatgcc caaaagaaag gctgcaggtc
      781 aaggtgatat gaggcaggag ccaaagagaa gatctgccag gttgtctgct atgcttgtgc
      841 cagttacacc agaggtgaag cctaaaagaa catcaagttc aaggaaaatg aagacaaaaa
      901 gtgatatgat ggaagaaaac atagatacaa gtgcccaagc agttgctgaa accaagcaag
      961 aagcagttgt tgaagaagac tacaatgaaa atgctaaaaa tggagaagcc aaaattacag
     1021 aggcaccagc ttctgaaaaa gaaattgtgg aagtaaaaga agaaaatatt gaagatgcca
     1081 cagaaaaggg aggagaaaag aaagaagcag tggcagcaga agtaaaaaat gaagaagaag
     1141 atcagaaaga agatgaagaa gatcaaaacg aagagaaagg ggaagctgga aaagaagaca
     1201 aagatgaaaa aggggaagaa gatggaaaag aggataaaaa tggaaatgag aaaggagaag
     1261 atgcaaaaga gaaagaagat ggaaaaaaag gtgaagacgg aaaaggaaat ggagaagatg
     1321 gaaaagagaa aggagaagat gaaaaagagg aagaagacag aaaagaaaca ggagatggaa
     1381 aagagaatga agatggaaaa gagaagggag ataaaaaaga ggggaaagat gtaaaagtca
     1441 aagaagatga aaaagagaga gaagatggaa aagaagatga aggtggaaat gaggaagaag
     1501 ctggaaaaga gaaagaagat ttaaaagaag aggaagaagg aaaagaggaa gatgagatca
     1561 aagaagatga tggaaaaaaa gaggagccac agagtattgt tggatccgac tacaaagacc
     1621 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     1681 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1741 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1801 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1861 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1921 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1981 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2041 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     2101 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2161 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2221 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2281 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     2341 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     2401 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     2461 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     2521 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     2581 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     2761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2941 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3001 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3121 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3181 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3241 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3301 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3361 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3601 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3961 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4261 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4381 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4501 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4561 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4621 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4681 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5161 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5401 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6721 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     6781 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     6841 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     6901 tgcatgaaga atctgcttag g
//
