LOCUS       V022128                 7925 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022128
VERSION     V022128
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..3415
                     /label="PRDM1(NM_001198)"
                     /note="PRDM1(NM_001198)"
                     /gene="PRDM1"
     CDS             3422..3487
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3525..3749
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3795..4223
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4237..4566
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4633..5427
                     /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    5601..5722
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5771..5787)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5795..5811
                     /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(5819..5849)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5864..5885
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6173..6758)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6929..7789)
                     /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(7790..7894)
                     /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 atgttggata tttgcttgga
      961 aaaacgtgtg ggtacgacct tggctgcccc caagtgtaac tccagcactg tgaggtttca
     1021 gggattggca gaggggacca aggggaccat gaaaatggac atggaggatg cggatatgac
     1081 tctgtggaca gaggctgagt ttgaagagaa gtgtacatac attgtgaacg accacccctg
     1141 ggattctggt gctgatggcg gtacttcggt tcaggcggag gcatccttac caaggaatct
     1201 gcttttcaag tatgccacca acagtgaaga ggttattgga gtgatgagta aagaatacat
     1261 accaaagggc acacgttttg gacccctaat aggtgaaatc tacaccaatg acacagttcc
     1321 taagaacgcc aacaggaaat atttttggag gatctattcc agaggggagc ttcaccactt
     1381 cattgacggc tttaatgaag agaaaagcaa ctggatgcgc tatgtgaatc cagcacactc
     1441 tccccgggag caaaacctgg ctgcgtgtca gaacgggatg aacatctact tctacaccat
     1501 taagcccatc cctgccaacc aggaacttct tgtgtggtat tgtcgggact ttgcagaaag
     1561 gcttcactac ccttatcccg gagagctgac aatgatgaat ctcacacaaa cacagagcag
     1621 tctaaagcaa ccgagcactg agaaaaatga actctgccca aagaatgtcc caaagagaga
     1681 gtacagcgtg aaagaaatcc taaaattgga ctccaacccc tccaaaggaa aggacctcta
     1741 ccgttctaac atttcacccc tcacatcaga aaaggacctc gatgacttta gaagacgtgg
     1801 gagccccgaa atgcccttct accctcgggt cgtttacccc atccgggccc ctctgccaga
     1861 agactttttg aaagcttccc tggcctacgg gatcgagaga cccacgtaca tcactcgctc
     1921 ccccattcca tcctccacca ctccaagccc ctctgcaaga agcagccccg accaaagcct
     1981 caagagctcc agccctcaca gcagccctgg gaatacggtg tcccctgtgg gccccggctc
     2041 tcaagagcac cgggactcct acgcttactt gaacgcgtcc tacggcacgg aaggtttggg
     2101 ctcctaccct ggctacgcac ccctgcccca cctcccgcca gctttcatcc cctcgtacaa
     2161 cgctcactac cccaagttcc tcttgccccc ctacggcatg aattgtaatg gcctgagcgc
     2221 tgtgagcagc atgaatggca tcaacaactt tggcctcttc ccgaggctgt gccctgtcta
     2281 cagcaatctc ctcggtgggg gcagcctgcc ccaccccatg ctcaacccca cttctctccc
     2341 gagctcgctg ccctcagatg gagcccggag gttgctccag ccggagcatc ccagggaggt
     2401 gcttgtcccg gcgccccaca gtgccttctc ctttaccggg gccgccgcca gcatgaagga
     2461 caaggcctgt agccccacaa gcgggtctcc cacggcggga acagccgcca cggcagaaca
     2521 tgtggtgcag cccaaagcta cctcagcagc gatggcagcc cccagcagcg acgaagccat
     2581 gaatctcatt aaaaacaaaa gaaacatgac cggctacaag acccttccct acccgctgaa
     2641 gaagcagaac ggcaagatca agtacgaatg caacgtttgc gccaagactt tcggccagct
     2701 ctccaatctg aaggtccacc tgagagtgca cagtggagaa cggcctttca aatgtcagac
     2761 ttgcaacaag ggctttactc agctcgccca cctgcagaaa cactacctgg tacacacggg
     2821 agaaaagcca catgaatgcc aggtctgcca caagagattt agcagcacca gcaatctcaa
     2881 gacccacctg cgactccatt ctggagagaa accataccaa tgcaaggtgt gccctgccaa
     2941 gttcacccag tttgtgcacc tgaaactgca caagcgtctg cacacccggg agcggcccca
     3001 caagtgctcc cagtgccaca agaactacat ccatctctgt agcctcaagg ttcacctgaa
     3061 agggaactgc gctgcggccc cggcgcctgg gctgcccttg gaagatctga cccgaatcaa
     3121 tgaagaaatc gagaagtttg acatcagtga caatgctgac cggctcgagg acgtggagga
     3181 tgacatcagt gtgatctctg tagtggagaa ggaaattctg gccgtggtca gaaaagagaa
     3241 agaagaaact ggcctgaaag tgtctttgca aagaaacatg gggaatggac tcctctcctc
     3301 agggtgcagc ctttatgagt catcagatct acccctcatg aagttgcctc ccagcaaccc
     3361 actacctctg gtacctgtaa aggtcaaaca agaaacagtt gaaccaatgg atcctctcga
     3421 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3481 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3541 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3601 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3661 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3721 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3781 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3841 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3901 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3961 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4021 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4081 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4141 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4201 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4321 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4381 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4441 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4501 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4561 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4801 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5161 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5401 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5461 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5581 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5641 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5701 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5761 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5821 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5881 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5941 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7921 acgtc
//
