LOCUS       V023958                 8363 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023958
VERSION     V023958
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             779..3043
                     /label="Dnm1l(NM_001360007)"
                     /note="Dnm1l(NM_001360007)"
                     /gene="Dnm1l"
     CDS             3050..3115
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3139..3727
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3793..3841
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4043..4471
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4485..4814
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4881..5675
                     /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    5851..5972
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6021..6037)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6045..6061
                     /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(6069..6099)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6114..6135
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6423..7011)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7182..8042)
                     /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(8043..8147)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggaggcgctg atcccggtca tcaataagct gcaggacgtc ttcaacacag tgggtgcgga
      841 catcatccag ctgcctcaga tcgtcgtagt gggaacgcag agcagtggga agagctcagt
      901 gctggaaagc ctagtgggca gggaccttct tcccagagga actggtgtgg tcacccggag
      961 acctctcatt ctgcagctag tccacgtttc accagaagat aaaagaaaaa caacaggaga
     1021 agaaaatggt aaatttcaga gctggaaccc tgccacatgg aaaaactcaa gacacctttc
     1081 caaaggagtt gaagcagaag aatggggtaa atttcttcac accaaaaaca agctttacac
     1141 agattttgat gaaattcgac aagaaattga aaatgaaact gaaagaattt caggaaataa
     1201 taagggggta agccctgagc caatccatct caaggttttc tcgcccaacg ttgtcaacct
     1261 gacacttgtg gatttaccgg gaatgaccaa agtacctgta ggcgatcagc ccaaggacat
     1321 cgagcttcag atcagagaac ttattcttcg gttcatcagt aatcccaatt ccattatcct
     1381 cgccgtcact gctgcaaata cagatatggc aacatcagaa gcactcaaga tttccagaga
     1441 ggtagatcca gatgggcgca gaactctagc tgtaatcact aaacttgatc tcatggatgc
     1501 gggtactgat gccatggatg tattgatggg aagggttatt ccagtcaagc ttggaataat
     1561 tggagtagtt aacagaagcc aactggatat taacaataag aagagtgtaa ctgattcaat
     1621 ccgtgatgag tatgcttttc ttcaaaagaa gtacccatct ctggccaaca gaaatggaac
     1681 aaagtatctt gctaggaccc tgaataggtt acttatgcat catatcagag attgtttacc
     1741 agagctgaaa acaagaataa atgtcttagc tgctcagtat cagtctcttc taaatagcta
     1801 tggtgaaccg gtggatgata aaagtgctac tttactccag cttattacca aatttgccac
     1861 agagtattgt aacacgattg aaggaaccgc aaagtacatt gaaacttctg agctatgcgg
     1921 tggtgctagg atttgttata ttttccatga gactttcggg cgaaccttag aatctgtgga
     1981 cccactaggt ggccttaaca ctattgacat cctgactgcc atcagaaatg ctactggccc
     2041 ccgtcctgct ttatttgtgc ctgaagtttc atttgagtta ctggtcaaac gtcagattaa
     2101 gcgtctagaa gagcccagcc tacggtgtgt ggagctggtc catgaggaga tgcagaggat
     2161 cattcagcat tgtagcaatt acagcacaca ggaattgtta cggttcccta aacttcacga
     2221 tgccatagtt gaagtagtga cctgtcttct tcgtaaaagg ttgcccgtga caaatgaaat
     2281 ggtgcataac ttagtggcaa ttgagctagc gtatatcaac acaaaacacc ccgactttgc
     2341 tgatgcctgt gggctaatga acaataatat agaggaacaa agaagaaaca ggctagcaag
     2401 agagctgcct tcagctggat cacgggacaa gtcttctaaa gttccaagtg ctttggcacc
     2461 tgcctcccag gagccccccc ctgctgcttc tgctgaggct gatggcaagt taattcagga
     2521 caacagaaga gaaactaaaa atgtcccatc tgcaggtggt gggattggag acggtggtca
     2581 ggaaccaaca acaggcaact ggagaggaat gctgaaaact tcaaaagctg aagaattact
     2641 tgctgaagaa aaatcaaaac caattccaat tatgccagca agtccacaga aaggccatgc
     2701 tgtcaatttg ctagatgtgc cagttccagt tgcaagaaaa ctgtctgccc gagaacagcg
     2761 agattgtgag gttattgaaa gacttatcaa atcatatttt ctaattgtca gaaagaatat
     2821 tcaagacagt gtcccaaagg cagtaatgca ttttttggtt aatcatgtga aagatactct
     2881 tcagagtgaa ctggtagggc agctgtataa gtcatcctta ttagatgacc ttctgactga
     2941 atccgaggac atggcccaac gaagaaaaga agcagcggat atgctgaagg cattacaagg
     3001 agccagtcaa attattgctg aaatccgaga gactcatctt tggctcgagg actacaaaga
     3061 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     3121 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3541 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3721 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3781 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3841 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3901 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3961 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     4021 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4081 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4141 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4201 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4261 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4321 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4381 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4441 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4741 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4801 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4861 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4921 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4981 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5041 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5101 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5161 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5221 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5281 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5341 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5401 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5461 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5521 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5581 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5641 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5701 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5761 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5821 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5881 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5941 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     6001 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6061 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6121 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6181 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6241 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6421 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6601 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6841 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7321 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7441 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7621 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7921 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8041 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8101 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8161 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     8221 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8281 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     8341 attgcatgaa gaatctgctt agg
//
