LOCUS       V020851                 7697 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020851
VERSION     V020851
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"
     CDS             683..2377
                     /label="USP21(NM_001014443)"
                     /note="USP21(NM_001014443)"
                     /gene="USP21"
     CDS             2384..2449
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2473..3061
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3127..3175
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3377..3805
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3819..4148
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4215..5009
                     /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    5185..5306
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5355..5371)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5379..5395
                     /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(5403..5433)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5448..5469
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5757..6345)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6516..7376)
                     /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(7377..7481)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgcccca ggcctctgag caccgcctgg gccgtacccg
      721 agagccacct gttaatatcc agccccgagt gggatccaag ctaccatttg cccccagggc
      781 ccgcagcaag gagcgcagaa acccagcctc tgggccaaac cccatgttac gacctctgcc
      841 tccccggcca ggtctgcctg atgaacggct caagaaactg gagctgggac ggggacggac
      901 ctcaggccct cgtcccagag gcccccttcg agcagatcat ggggttcccc tgcctggctc
      961 accaccccca acagtggctt tgcctctccc atctcggacc aacttagccc gttccaagtc
     1021 tgtgagcagt ggggacttgc gtccaatggg gattgccttg ggagggcacc gtggcaccgg
     1081 agagcttggg gctgcactga gccgcttggc cctccggcct gagccaccca ctttgagacg
     1141 tagcacttct ctccgccgcc tagggggctt tcctggaccc cctaccctgt tcagcatacg
     1201 gacagagccc cctgcttccc atggctcctt ccacatgata tccgcccggt cctctgagcc
     1261 tttctactct gatgacaaga tggctcatca cacactcctt ctgggctctg gtcatgttgg
     1321 ccttcgaaac ctgggaaaca cgtgcttcct gaatgctgtg ctgcagtgtc tgagcagcac
     1381 tcgacctctt cgggacttct gtctgagaag ggacttccgg caagaggtgc ctggaggagg
     1441 ccgagcccaa gagctcactg aagcctttgc agatgtgatt ggtgccctct ggcaccctga
     1501 ctcctgcgaa gctgtgaatc ctactcgatt ccgagctgtc ttccagaaat atgttccctc
     1561 cttctctgga tacagccagc aggatgccca agagttcctg aagctcctca tggagcggct
     1621 acaccttgaa atcaaccgcc gaggccgccg ggctccaccg atacttgcca atggtccagt
     1681 tccctctcca ccccgccgag gaggggctct gctagaagaa cctgagttaa gtgatgatga
     1741 ccgagccaac ctaatgtgga aacgttacct ggagcgagag gacagcaaga ttgtggacct
     1801 gtttgtgggc cagttgaaaa gttgtctcaa gtgccaggcc tgtgggtatc gctccacgac
     1861 cttcgaggtt ttttgtgacc tgtccctgcc catccccaag aaaggatttg ctgggggcaa
     1921 ggtgtctctg cgggattgtt tcaacctttt cactaaggaa gaagagctag agtcggagaa
     1981 tgccccagtg tgtgaccgat gtcggcagaa aactcgaagt accaaaaagt tgacagtaca
     2041 aagattccct cgaatcctcg tgctccatct gaatcgattt tctgcctccc gaggctccat
     2101 caaaaaaagt tcagtaggtg tagactttcc actgcagcga ctgagcctag gggactttgc
     2161 cagtgacaaa gccggaagtc ctgtatacca gctgtatgcc ctttgcaacc actcaggcag
     2221 cgtccactat ggccactaca cagccctgtg ccggtgccag actggttggc atgtctacaa
     2281 tgactctcgt gtctcccctg tcagtgaaaa ccaggtggca tccagcgagg gctacgtgct
     2341 gttctaccaa ctgatgcagg agccaccccg gtgcctgctc gaggactaca aagaccatga
     2401 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2461 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2521 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2581 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2641 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2701 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2761 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2821 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2881 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2941 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3001 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3061 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3121 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     3181 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3241 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3301 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3721 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4081 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4141 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4201 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4981 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     5041 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5101 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5161 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5221 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5281 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5341 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5941 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6181 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7501 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     7561 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7621 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     7681 tgaagaatct gcttagg
//
