LOCUS       V044669                 9186 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044669
VERSION     V044669
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..3866
                     /label="Senp7(NM_025483)"
                     /note="Senp7(NM_025483)"
                     /gene="Senp7"
     CDS             3873..3938
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3962..4550
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4616..4664
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4866..5294
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5308..5637
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5704..6498
                     /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    6674..6795
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6844..6860)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6868..6884
                     /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(6892..6922)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6937..6958
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7246..7834)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8005..8865)
                     /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(8866..8970)
                     /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 ccaccatgga cagggccagg cctgggcgtc
      781 ggcgagcttc atcagagatt gttacagagg gaaaacggaa aaagtcatca ccagctgatt
      841 tacagaagat aacaaagttg ttaactgtaa agtcagagga tgttcttgct cagtcaccat
      901 tgtccaaact cagaggctca gaatgctggt ggacaagaag cctaagaaat aaagtcatct
      961 gtctagacca caaaaaacca aaagctgccc gtgggtgtcc tcctaaggga ttaccaaaaa
     1021 ggcatctcag agttatgttg acgaatgttc tatggacgga cttaggacga gaattcagaa
     1081 agaccctgcc tagaaaggat gctaatttat gtgctcccag caaggtgcaa tcagactcat
     1141 tgccttcgac atctgttgac agcatagaga catgtcaaag attagatcct cttcaccaaa
     1201 gccttaattt atccgaaagg acacccagag ttatactgac ggatatccgg caaacagaat
     1261 taggaagaaa atatttaaag atcccacctg taactgaggc cagtttgagt gatacagcca
     1321 acctgaaatc agagcaactt tcttcatcat ctgatggcag cttagagtct tgtcagagtg
     1381 taaatcatca caagagcttt ttatctgaaa gtggtcccaa accaagtagg acaggtgacg
     1441 ttcctgcaaa ggaggctgca tgtgggggac agaagcaggg tgatgatgga ggagtcactc
     1501 ctgagatggc tgctcctcat cctaaagact ttaatactgg aaataaaggt tgtgattatc
     1561 ttgaagaggg gaccagcaac aagaatactt catattctta ttcggaaatg gaccatactc
     1621 cagtttccag gaagagaaag aaaaggggta gaagtaattt tcatgattct cataattcta
     1681 agacttcttt ggataagcca acagaacata caaaagaaga agaaaatgac tcttcagtct
     1741 cccgtaagct tgaagagtcc ggtgaagact ctcaccagga cccagcacca cccgaaggac
     1801 tcgcgccaga atctttagag agcgaggcca ctaacctgcg ctcattcgct gctggtcagg
     1861 agccagatgc tagcgctgcc tctggaaggg cctcttcacc caacaaaagc ttggagagct
     1921 ctgcttctag tgaagtgtcg gagaattctt ccgtggctgt gaagggtgag gcgttgactc
     1981 taaaagaagc atcccctccg ggtggaagca gtgaagagag ccagttgttg atctcagctg
     2041 agccaattgt agtttccagt gatgaagaag gaccggttga acataagaat tcagtaattc
     2101 taaaattaca gcctcctcat gagattatga gtgaaaatca gggcacttct gacccacagt
     2161 tgtcagagct gacactcggt gcatgtgaat ctgtacaggt aacatctgaa ttatttccat
     2221 acaatcctga tgtagaaaac atttcctgta ttaaatctaa tagtgaaatg gatctgaaat
     2281 tggatttcat atttacatgt gtgtatattg gtaaaataaa aggaactcca aaaggttgtg
     2341 tcacgttcac aaagaagtat attaaaatcc cgtttcaagt gtccacaaat gagatttccc
     2401 tgacagtgga caccgcacgt ttaaagaggt tcgggttatg ggaaagcaag gatgaggacc
     2461 acagcaagag gagccacgct atccttttcc tctggctttc ctcagactac cttcaagaca
     2521 ttcagaccca gctagaaaac cccatgctaa gccagcaatc aaaagccaat gaatttattt
     2581 tccttgagtt aaacagttcc atttcacaaa gagaagaatt gaaattgaaa gatattatga
     2641 tggaaatcag tacaaaaaat ggaaatttac acctgtcttg tccattgccc tgggtccagg
     2701 cacttccttt gtttcaggat ctctctccac aagaaatctc ttttcttcac tattactatg
     2761 cttcagcttc tgctctcccc actgctgctg gggcagacat gaagaagaag tcagtgtctc
     2821 agccctcaaa ttcagacacc atcaagccta cttacacctt cctacataaa caaagtagtg
     2881 gttgctactc gctctctatt acttcaagtc cagaggagga atggcaggaa gtcaggaaca
     2941 cgggacccgt tcagaagttg attgtttatc ctccaccacc tactaaggga ggcttgggag
     3001 taactaatga agatctagag tgtttagaag aaggagaatt tcttaacgat gtaatcattg
     3061 acttttacct taagtatctt ctattggaga aggcatcaga tgaacttgtt gaacgaagtc
     3121 acatttttag tagctttttc tataaatgct tgacaagaaa ggaaaataat ttaacagaag
     3181 ataatccaga tctatcagtg gctcagagga gacaccgaag agtgagaaca tggactcgtc
     3241 atataaacat ttttaataaa gactacatct ttgtacctgt taatgaatcg tctcactggt
     3301 atcttgcagt catttgtttc ccatggctag aagaggctgt gtatgaagat tgtccacaaa
     3361 ctgtgtccca acagttccag ggtcagcagt ctcagcatga ccacaaaatg acagataatg
     3421 atccacatac cacttcaact gtgtccacga gtgcagagga ttcccaaagt accgaggtga
     3481 atatgtcagt accaaagaaa atgtgtaagc ggccatgtat tctcatacta gactccctga
     3541 aagctgcttc tatacaaaac acagttcaga acttacgaga gtatttggag gtagagtggg
     3601 aggtgaaacg aaaaacgcac cgtgaattca gtaaaaccaa tatggtcgac ctgtgcccca
     3661 aagtccctaa gcaggataac agcagtgact gtggagtcta tttgttgcag tatgtggaga
     3721 gctttttcca ggatcccatt gttaactttg aacttccaat tcacttggag aaatggtttc
     3781 ctcgccatgt aataaagacc aaacgggaag atattcgaga gctcattttg aaacttcact
     3841 tacagcaaca gaagggcggc agctgcggat ccgactacaa agaccatgac ggtgattata
     3901 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3961 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4021 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4081 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4141 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4201 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4261 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4321 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4381 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4441 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4501 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     4561 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4621 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4681 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4741 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4801 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4861 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     5041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5221 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5281 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5341 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5401 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5461 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5521 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5581 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5641 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5701 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5761 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5821 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5881 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5941 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6001 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6061 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6121 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6181 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6241 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6301 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6361 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6421 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6481 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     6541 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6601 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6661 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6781 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6841 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7021 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7081 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7141 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7201 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7261 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7321 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7381 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7441 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7501 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7561 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7621 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7681 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7741 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7801 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7861 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7921 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7981 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8041 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8101 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8161 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8221 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8281 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8341 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8401 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8461 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8521 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8581 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8641 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8701 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8761 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8821 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8881 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8941 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9001 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     9061 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9181 cttagg
//
