LOCUS       V045896                 9411 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045896
VERSION     V045896
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..4091
                     /label="SENP6(NM_015571)"
                     /note="SENP6(NM_015571)"
                     /gene="SENP6"
     CDS             4098..4163
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4187..4775
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4841..4889
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5091..5519
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5533..5862
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5929..6723
                     /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    6899..7020
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7069..7085)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7093..7109
                     /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(7117..7147)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7162..7183
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7471..8059)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8230..9090)
                     /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(9091..9195)
                     /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 ccaccatggc ggccggcaag agcggcggta
      781 gcgcagggga gattactttt ctggaagctt tggctagatc agagtctaag agagatggag
      841 gttttaaaaa taattggagc tttgatcatg aagaagaaag tgaaggagat acagataaag
      901 atgggacaaa tctgctcagt gtggatgaag atgaggattc tgaaacctca aaaggaaaaa
      961 agttaaatcg tcgatctgaa attgttgcta atagctctgg tgaattcatc ttgaagacat
     1021 atgtaagacg aaacaagtct gaaagtttta aaactttgaa aggcaaccca attggactta
     1081 acatgttgag caacaataag aaattgagtg aaaatacgca aaatacgtca ttatgttctg
     1141 gaactgtagt tcatggtaga cgttttcatc atgctcatgc acagatacca gtagtaaaaa
     1201 cagcagccca aagcagtctg gaccgaaaag aaaggaaaga atacccacct catgtccaaa
     1261 aagttgaaat taatcctgta aggttaagtc ggctccaagg tgttgaacgt ataatgaaga
     1321 aaacagaaga gtccgaatca caagtggagc ctgaaattaa gaggaaagta caacagaaac
     1381 gacactgtag tacctatcag cctactcctc ctctatctcc tgcttcaaaa aaatgtttaa
     1441 cccatttaga ggatttgcaa agaaattgca gacaagctat tactttgaat gagtctactg
     1501 gaccattatt aagaacgtca attcatcaga attctggagg acagaagtca caaaacacag
     1561 gattaacaac caagaagttt tatggcaaca atgtggaaaa ggttccaatt gatattattg
     1621 tgaattgtga tgacagtaaa cacacttatt tacagactaa tggaaaagtc attttacctg
     1681 gggcaaaaat acccaaaatc acaaacttga aagaaaggaa aacaagtttg tcagacctaa
     1741 atgatccaat cattttgtcc agtgatgatg atgatgacaa cgacagaact aacagaagag
     1801 aaagcatatc tcctcagcct gctgattcag catgttcttc ccctgcacca tccactggaa
     1861 aagtagaagc agcgctaaat gaaaatactt gcagagcaga gcgtgaacta cgaagcattc
     1921 cagaagactc agagttaaat acagttacat tgccaagaaa agcaagaatg aaagaccagt
     1981 ttggcaattc tattatcaac acacctctga aacgtcgtaa agtgttttct caagaacctc
     2041 cagatgcttt agctttaagc tgccaaagtt cctttgacag tgtcatttta aactgtcgaa
     2101 gtatacgagt aggaacactc ttccggctgt taatagagcc tgtaattttt tgtttagatt
     2161 ttatcaagat acagctagac gaaccagacc atgatcctgt agagattata ttaaatacct
     2221 ctgatctaac taaatgtgaa tggtgtaatg tccgaaaatt acctgtagtg tttcttcaag
     2281 caattccagc agtttatcaa aagctgagca tccaactgca aatgaataag gaggataaag
     2341 tttggaatga ttgtaaagga gtaaataaat taacaaattt agaagaacaa tatataattt
     2401 taatttttca aaatggcctt gatcctccgg caaatatggt atttgaaagt atcattaatg
     2461 aaattggtat aaagaataac atctccaatt tttttgcgaa aattcccttt gaagaagcta
     2521 atggcagact tgttgcctgt acaagaacct atgaagagag catcaaagga agttgtgggc
     2581 aaaaggaaaa caaaattaaa actgtatcat ttgaatctaa aatacaactt agaagcaaac
     2641 aagaatttca gttttttgat gaagaagaag aaactggaga aaaccacacc atcttcattg
     2701 gcccagtaga aaagttgata gtatatccac cacctccagc taagggaggc atctctgtta
     2761 ccaatgagga cctgcactgt ctaaatgaag gagaattttt aaatgatgtt attatagact
     2821 tttatttgaa atacttggtg cttgaaaaac tgaagaagga agacgctgac cgaattcata
     2881 tattcagttc ttttttctat aaacgcctta atcagagaga gaggagaaat catgaaacaa
     2941 ctaatctgtc aatacagcaa aaacggcatg ggagagtaaa aacatggacc cggcacgtag
     3001 atatttttga gaaggatttt atttttgtac cccttaatga agctgcacac tggtttttgg
     3061 ctgttgtttg tttccccggt ttggaaaaac caaagtatga acctaatcct cattaccatg
     3121 aaaatgctgt catacagaaa tgttcaactg tagaggacag ttgtatttct tcttcagcca
     3181 gtgaaatgga gagttgttca caaaactctt ctgccaagcc tgtaattaag aagatgctaa
     3241 acaaaaaaca ttgcatagct gtaattgatt ccaatcctgg gcaggaagaa agtgaccctc
     3301 gttataagag aaacatatgc agtgtaaaat acagtgtgaa aaaaataaat catactgcga
     3361 gtgaaaatga agaattcaat aaaggagaat ctacatccca gaaagttgct gataggacta
     3421 aaagtgagaa tggcctacag aatgaaagtt taagttccac acatcataca gatggcttaa
     3481 gcaaaatcag actaaactat agcgatgaat cacctgaagc tggtaaaatg cttgaagatg
     3541 aactcgtcga cttctcagaa gatcaggata accaggatga tagcagtgac gatggattcc
     3601 tcgctgatga caactgcagt tcagaaatag gacagtggca tttaaagcct actatctgta
     3661 aacaaccttg tatcctactt atggactcac tccgaggccc ttctcggtca aatgttgtca
     3721 aaattttaag agagtattta gaagtggaat gggaagttaa aaaaggaagc aaaagaagtt
     3781 tttccaaaga tgttatgaag ggctctaatc caaaagtacc acagcaaaac aacttcagtg
     3841 actgtggtgt atatgtattg cagtatgtag agagcttttt tgagaatcca attctcagtt
     3901 ttgaactacc tatgaatttg gcaaactggt ttcctccacc aagaatgaga acaaaaagag
     3961 aagaaatccg aaacataatt ctgaagctac aggaagatca gagcaaagag aaaagaaagc
     4021 ataaggacac ttactcaaca gaagcacctt taggcgaagg aacagaacaa tatgtcaata
     4081 gtatctcaga tggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4141 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     4201 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4261 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4321 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4381 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4441 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4501 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4561 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4621 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4681 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4741 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     4801 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4861 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4921 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4981 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5041 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     5101 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5161 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5221 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     5281 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5341 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5401 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5461 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5521 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5581 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5701 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5761 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5821 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5881 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5941 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6001 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6061 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6121 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6181 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6241 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6301 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6361 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6421 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6481 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6541 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6601 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6661 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6721 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6781 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6841 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6901 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6961 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7021 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     7081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7141 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     7681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9241 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     9301 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     9361 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
