LOCUS       V021095                 8750 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021095
VERSION     V021095
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..4204
                     /label="SMC6(NM_024624)"
                     /note="SMC6(NM_024624)"
                     /gene="SMC6"
     CDS             4235..4264
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4280..4297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4347..4571
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4617..5045
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5059..5388
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5455..6249
                     /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    6423..6544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6593..6609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6617..6633
                     /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(6641..6671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6686..6707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6995..7583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7754..8614)
                     /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(8615..8719)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagccac catggccaaa agaaaggaag aaaatttttc
      961 ctctcctaaa aatgccaaaa ggccaagaca agaagaattg gaggattttg ataaagatgg
     1021 tgacgaagac gaatgtaaag gtactacttt gactgcagca gaagttggaa taattgagag
     1081 tattcaccta aaaaacttca tgtgtcattc aatgcttgga ccttttaagt ttggttctaa
     1141 tgtcaacttt gttgttggca acaatggaag tgggaagagt gcagtactca cagctctcat
     1201 agtcggtctt ggtggaagag cagttgctac taatagagga tcctctttaa aaggttttgt
     1261 gaaagatgga cagaactctg cagatatctc aataacattg aggaacagag gagatgatgc
     1321 ctttaaagcc agtgtgtatg gtaactctat acttatacag caacacatca gcatagatgg
     1381 aagtcgatct tataaactta aaagtgcaac aggctccgtg gtttccacga ggaaagaaga
     1441 gctgattgca attcttgatc attttaacat ccaggtggat aatccagttt ctgttttaac
     1501 acaagaaatg agcaagcagt tcttacagtc taaaaatgaa ggagacaaat acaaattctt
     1561 catgaaagca acgcaacttg aacagatgaa ggaagattat tcatacatta tggaaacgaa
     1621 agaaagaaca aaggagcaga tacatcaagg agaagagcgg cttactgaac taaagcgcca
     1681 gtgtgtagag aaagaggaac gttttcaaag tattgctggt ttaagtacaa tgaagactaa
     1741 tttagagtcc ttgaaacatg aaatggcttg ggcagtggtc aatgaaattg aaaaacaatt
     1801 gaatgccatc agagataata tcaaaattgg agaagatcgt gctgctagac ttgacaggaa
     1861 aatggaagaa cagcaggtca gacttaatga ggcagaacaa aagtacaagg atattcaaga
     1921 caaactagaa aagattagtg aagagacaaa tgcacgagca ccagaatgta tggcattgaa
     1981 agcagatgtt gttgctaaga aaagggccta taatgaagct gaggttttat ataaccgatc
     2041 cttaaacgaa tataaagcat taaagaaaga tgatgagcag ctttgtaaac gaattgaaga
     2101 gctgaaaaaa agtactgacc aatctttgga acctgaacgg ttggaaagac aaaaaaaaat
     2161 atcttggtta aaagagagag taaaggcctt tcaaaatcaa gaaaattcag tcaatcaaga
     2221 gatcgaacag tttcagcaag ccatagaaaa ggacaaagaa gaacatggca aaattaagag
     2281 agaagaatta gatgtgaagc atgcactgag ctacaatcag aggcaactga aagaattgaa
     2341 agatagtaaa actgatcgac tcaaaagatt tggccctaat gttccagctc ttcttgaagc
     2401 catagatgat gcttatagac aaggacattt tacctataaa cctgtaggcc ctttaggagc
     2461 ttgcattcat cttcgggacc cagaacttgc tttggctatt gaatcttgct taaaagggct
     2521 tctgcaggcc tattgttgcc ataatcatgc tgatgaaagg gtccttcagg cactcatgaa
     2581 aaggttttat ttaccaggga cctcacggcc accgataata gtttctgagt ttcggaatga
     2641 gatatatgat gtaagacaca gagctgctta tcatccagac tttccaacag ttctgacagc
     2701 tttagaaata gataatgcgg ttgtggcaaa tagcctaatt gacatgagag gcatagagac
     2761 agtgctacta atcaaaaata attctgtagc tcgtgcagta atgcagtccc aaaagccacc
     2821 caaaaattgt agagaagctt ttactgctga tggtgatcaa gtttttgcag gacgttatta
     2881 ttcatctgaa aatacaagac ctaagttcct aagcagagat gtggattctg aaataagtga
     2941 cttggagaat gaggttgaaa ataagacggc ccagatatta aatcttcagc aacatttatc
     3001 tgcccttgaa aaagatatta aacacaatga ggaacttctt aaaaggtgcc aactacatta
     3061 taaagaacta aagatgaaaa taagaaaaaa tatttctgaa attcgggaac ttgagaacat
     3121 agaagaacac cagtctgtag atattgcaac tttggaagat gaagctcagg aaaataaaag
     3181 caaaatgaaa atggttgagg aacatatgga gcaacaaaaa gaaaatatgg agcatcttaa
     3241 aagtctgaaa atagaagcag aaaataagta tgatgcaatt aaattcaaaa ttaatcaact
     3301 atcggagcta gcagacccac ttaaggatga attaaacctt gctgattctg aagtggataa
     3361 ccaaaaacga gggaaacgac attatgaaga aaaacaaaaa gaacacttgg ataccttaaa
     3421 taaaaagaaa cgagaactgg atatgaaaga gaaagaacta gaggagaaaa tgtcacaagc
     3481 aagacaaatc tgcccagagc gtatagaagt agaaaaatct gcatcaattc tggacaaaga
     3541 aattaatcga ttaaggcaga agatacaggc agaacatgct agtcatggag atcgagagga
     3601 aataatgagg cagtaccaag aagcaagaga gacctatctt gatctggata gtaaagtgag
     3661 gactttaaaa aagtttatta aattactggg agaaatcatg gagcacagat tcaagacata
     3721 tcaacaattt agaaggtgtt tgactttacg atgcaaatta tactttgaca acttactatc
     3781 tcagcgggcc tattgtggaa aaatgaattt tgaccacaag aatgaaactc taagtatatc
     3841 agttcagcct ggagaaggaa ataaagctgc tttcaatgac atgagagcct tgtctggagg
     3901 tgaacgttct ttctccacag tgtgttttat tctttccctg tggtccatcg cagaatctcc
     3961 tttcagatgc ctggatgaat ttgatgtcta catggatatg gttaatagga gaattgccat
     4021 ggacttgata ctgaagatgg cagattccca gcgttttaga cagtttatct tgctcacacc
     4081 tcaaagcatg agttcacttc catccagtaa actgataaga attctccgaa tgtctgatcc
     4141 tgaaagagga caaactacat tgcctttcag acctgtgact caagaagaag atgatgacca
     4201 aaggggatcc gagctcggta ccaagcttgg gcccgaacaa aaactcatct cagaagagga
     4261 tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt
     4321 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4381 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4441 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4501 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     4621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5041 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5401 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
