LOCUS       V022612                 7805 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022612
VERSION     V022612
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             941..3295
                     /label="SP1(NM_138473)"
                     /note="SP1(NM_138473)"
                     /gene="SP1"
     CDS             3302..3367
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3405..3629
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3675..4103
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4117..4446
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4513..5307
                     /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    5481..5602
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5651..5667)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5675..5691
                     /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(5699..5729)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5744..5765
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6053..6638)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6809..7669)
                     /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(7670..7774)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagcgacc aagatcactc
      961 catggatgaa atgacagctg tggtgaaaat tgaaaaagga gttggtggca ataatggggg
     1021 caatggtaat ggtggtggtg ccttttcaca ggctcgaagt agcagcacag gcagtagcag
     1081 cagcactgga ggaggagggc aggagtccca gccatcccct ttggctctgc tggcagcaac
     1141 ttgcagcaga attgagtcac ccaatgagaa cagcaacaac tcccagggcc cgagtcagtc
     1201 agggggaaca ggtgagcttg acctcacagc cacacaactt tcacagggtg ccaatggctg
     1261 gcagatcatc tcttcctcct ctggggctac ccctacctca aaggaacaga gtggcagcag
     1321 taccaatggc agcaatggca gtgagtcttc caagaatcgc acagtctctg gtgggcagta
     1381 tgttgtggct gccgctccca acttacagaa ccagcaagtt ctgacaggac tacctggagt
     1441 gatgcctaat attcagtatc aagtaatccc acagttccag accgttgatg ggcaacagct
     1501 gcagtttgct gccactgggg cccaagtgca gcaggatggt tctggtcaaa tacagatcat
     1561 accaggtgca aaccaacaga ttatcacaaa tcgaggaagt ggaggcaaca tcattgctgc
     1621 tatgccaaac ctactccagc aggctgtccc cctccaaggc ctggctaata atgtactctc
     1681 aggacagact cagtatgtga ccaatgtacc agtggccctg aatgggaaca tcaccttgct
     1741 acctgtcaac agcgtttctg cagctacctt gactcccagc tctcaggcag tcacgatcag
     1801 cagctctggg tcccaggaga gtggctcaca gcctgtcacc tcagggacta ccatcagttc
     1861 tgccagcttg gtatcatcac aagccagttc cagctccttt ttcaccaatg ccaatagcta
     1921 ctcaactact actaccacca gcaacatggg aattatgaac tttactacca gtggatcatc
     1981 agggaccaac tctcaaggcc agacacccca gagggtcagt gggctacagg ggtctgatgc
     2041 tctgaacatc cagcaaaacc agacatctgg aggctcattg caagcaggcc agcaaaaaga
     2101 aggagagcaa aaccagcaga cacagcagca acaaattctt atccagcctc agctagttca
     2161 agggggacag gccctccagg ccctccaagc agcaccattg tcagggcaga cctttacaac
     2221 tcaagccatc tcccaggaaa ccctccagaa cctccagctt caggctgttc caaactctgg
     2281 tcccatcatc atccggacac caacagtggg gcccaatgga caggtcagtt ggcagactct
     2341 acagctgcag aacctccaag ttcagaaccc acaagcccaa acaatcacct tagccccaat
     2401 gcagggtgtt tccttggggc agaccagcag cagcaacacc actctcacac ccattgcctc
     2461 agctgcttcc attcctgctg gcacagtcac tgtgaatgct gctcaactct cctccatgcc
     2521 aggcctccag accattaacc tcagtgcatt gggtacttca ggaatccagg tgcacccaat
     2581 tcaaggcctg ccgttggcta tagcaaatgc cccaggtgat catggagctc agcttggtct
     2641 ccatggggct ggtggtgatg gaatacatga tgacacagca ggtggagagg aaggagaaaa
     2701 cagcccagat gcccaacccc aagccggtcg gaggacccgg cgggaagcat gcacctgccc
     2761 ctactgtaaa gacagtgaag gaaggggctc gggggatcct ggcaaaaaga aacagcatat
     2821 ttgccacatc caaggctgtg ggaaagtgta tggcaagacc tctcacctgc gggcacactt
     2881 gcgctggcat acaggcgaga ggccatttat gtgtacctgg tcatactgtg ggaaacgctt
     2941 cacacgttcg gatgagctac agaggcacaa acgtacacac acaggtgaga agaaatttgc
     3001 ctgccctgag tgtcctaagc gcttcatgag gagtgaccac ctgtcaaaac atatcaagac
     3061 ccaccagaat aagaagggag gcccaggtgt agctctgagt gtgggcactt tgcccctgga
     3121 cagtggggca ggttcagaag gcagtggcac tgccactcct tcagccctta ttaccaccaa
     3181 tatggtagcc atggaggcca tctgtccaga gggcattgcc cgtcttgcca acagtggcat
     3241 caacgtcatg caggtggcag atctgcagtc cattaatatc agtggcaatg gcttcctcga
     3301 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3361 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3601 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3661 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4081 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4201 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4261 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4321 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4381 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4441 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4501 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4561 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4621 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4681 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4741 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4801 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4861 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4921 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4981 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5041 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5101 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5161 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5221 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5281 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5341 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5401 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5461 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5521 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5581 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5641 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5701 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5761 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5821 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5881 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5941 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7801 acgtc
//
