LOCUS       V020827                 7529 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020827
VERSION     V020827
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..2209
                     /label="SPAG6(NM_012443)"
                     /note="SPAG6(NM_012443)"
                     /gene="SPAG6"
     CDS             2216..2281
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2305..2893
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2959..3007
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3209..3637
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3651..3980
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4047..4841
                     /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    5017..5138
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5187..5203)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5211..5227
                     /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(5235..5265)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5280..5301
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5589..6177)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6348..7208)
                     /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(7209..7313)
                     /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 ccatgagtca gaggcaggtg ctgcaagtgt tcgagcaata
      721 ccagaaggcc aggacccagt tcgtgcagat ggtggcggag ctggcgacta gaccccaaaa
      781 catcgagacg ctgcagaacg cgggtgtaat gtctttgctg agaactcttc ttctggacgt
      841 ggtcccaaca attcaacaga ctgctgcttt ggctcttggg agactggcca attataatga
      901 tgacctagca gaagctgttg tgaagtgcga cattcttcca cagcttgttt attcattggc
      961 agaacagaat cgcttctaca agaaagcagc tgcctttgtg ttacgagcag ttggtaaaca
     1021 ttctccccag ctagctcagg caatagtcga ttgtggagca ctggatacgc tggtcatatg
     1081 cttggaagat tttgaccctg gagtcaagga ggctgcagcc tgggcactta gatatattgc
     1141 aagacataat gcagaactgt cacaagctgt ggtggatgca ggagctgttc ctcttttagt
     1201 actctgtatc caggagccag aaattgcttt gaaaaggatt gctgcttcgg ccctcagtga
     1261 tattgcaaag cattctccag agttagcaca gacagtagtg gatgcaggag ctgttgctca
     1321 tttagcccag atgatcctga accctgatgc taaattgaag catcagatcc tttcagctct
     1381 cagtcaggtt tcaaaacatt ccgtggatct ggcagaaatg gttgttgaag cagagatttt
     1441 tccagttgta cttacctgtc tgaaggacaa ggatgaatac gtgaagaaaa atgcttctac
     1501 tttaattaga gagattgcaa aacatacacc cgagctttca cagctggtag ttaacgcagg
     1561 aggggttgct gccgtgattg actgcattgg gtcctgcaaa gggaacacac ggctgcctgg
     1621 catcatgatg cttggttatg tagcagctca ttctgagaac ctagcaatgg cagtcatcat
     1681 ttctaagggt gtaccccagt tgtcagtctg cttgtcagaa gaaccggaag atcatattaa
     1741 ggctgcagct gcttgggcct taggacagat tggaagacac actcctgaac acgcacgggc
     1801 tgttgcagtc acaaatactt tgccagttct gctttctttg tacatgtcaa cagaaagttc
     1861 tgaggatctc caagtaaaaa gtaaaaaagc cataaagaat atcctgcaaa aatgtaccta
     1921 cttaccagcc cttgaaccat ttctatatga tgctcctccc aatattctga aacatgtggt
     1981 tggacagttc agtaaggtgc tgccgcatga tagcaaagct cgacgacttt ttgtaacaag
     2041 tggtggcctt aaaaaagttc aagagataaa agcagaacct ggttctctcc ttcaagaata
     2101 catcaacagt attaacagtt gttaccccga ggaaatagtg aggtattatt cccctggata
     2161 ttcagataca cttctgcaga gggtggacag ctatcaacca cttaataacc tcgaggacta
     2221 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2281 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     2341 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2401 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2461 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2521 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2581 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2641 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2701 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2761 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2821 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2881 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     2941 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3001 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3061 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3121 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     3181 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3241 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3301 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3361 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3421 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3481 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3541 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3601 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3661 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3901 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3961 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4021 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4081 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4141 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4201 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4261 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4321 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4381 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4441 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4501 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4561 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4621 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4681 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4741 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4801 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4861 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4921 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4981 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5041 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5101 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5161 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5221 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5281 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5341 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5401 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5461 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5521 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5761 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6001 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7321 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     7381 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7441 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7501 ccgacaattg catgaagaat ctgcttagg
//
