LOCUS       V029660                 7776 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029660
VERSION     V029660
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..2456
                     /label="PHF1(NM_024165)"
                     /note="PHF1(NM_024165)"
                     /gene="PHF1"
     CDS             2463..2528
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2552..3140
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3206..3254
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3456..3884
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3898..4227
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4294..5088
                     /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    5264..5385
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5434..5450)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5458..5474
                     /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(5482..5512)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5527..5548
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5836..6424)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6595..7455)
                     /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(7456..7560)
                     /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 gcagcccccc cggctgagcc
      781 gctctggtgc ctcctcactt tgggacccag cttctcctgc tcccacctct ggccccaggc
      841 ctcggctttg ggagggtcaa gatgtgctgg ccagatggac tgatgggctg ctatacttgg
      901 gtaccatcaa aaaggtggac agtgctaggg aggtgtgtct ggtccagttt gaggatgatt
      961 cgcagtttct ggttctatgg aaagacatta gccctgctgc cctccctgga gaggaactcc
     1021 tctgttgtgt ctgtcgctct gagactgtgg tccctgggaa ccggctggtc agctgtgaga
     1081 agtgtcgcca tgcttatcac caggactgcc atgttcccag ggctccagcc cctggagagg
     1141 gagagggcac atcctgggta tgccgccagt gtgtctttgc gatcgccacc aagaggggag
     1201 gtgccctgaa gaagggcccc tatgcccggg ccatgctggg tatgaagctt tctctgccat
     1261 atggactgaa ggggctggac tgggatgctg gacatctgag caaccgacag cagagttact
     1321 gttactgtgg tggccctggg gagtggaacc tgaaaatgct gcagtgccgg agctgcctgc
     1381 agtggttcca tgaggcctgc acccagtgtc tgagcaagcc cctcctctat ggggacaggt
     1441 tctatgaatt tgaatgctgt gtgtgtcgcg ggggccctga gaaagtccgg agactacagc
     1501 ttcgctgggt ggatgtggcc catcttgtcc tgtatcacct cagtgtttgc tgtaagaaga
     1561 aatactttga ttttgatcgt gagatcctcc ccttcacttc tgagaattgg gacagtttgc
     1621 tcctggggga gctttcagac acccccaaag gagaacgttc ttccaggctc ctctctgctc
     1681 ttaacagcca caaggaccgt ttcatttcag ggagagagat taagaagagg aaatgtttgt
     1741 ttggtctcca tgctcggatg cctccccctg tggagccccc tactggagat ggagcactca
     1801 ccagcttccc ttcagggcag ggccctgggg gaggggtctc acgtcccctg gggaagcgcc
     1861 ggaggccgga gccagagccc ctgaggagga ggcagaaggg gaaagtggag gagctggggc
     1921 caccctcagc agtgcgcaat cagcccgagc cccaggagca gagggagcgg gctcatctgc
     1981 agagggcact gcaggcctca gtgtctccac catcccccag ccctaaccag agttaccagg
     2041 gcagcagcgg ctacaacttc cggcccacag atgcccgctg cctgcccagc agccccatcc
     2101 ggatgtttgc ttccttccac ccttctgcca gcaccgcagg gacctctggg gacagtggac
     2161 ccccagacag gtcacccctg gaacttcaca ttggtttccc cacagacatc cctaaaagtg
     2221 ccccccactc gatgactgcc tcatcttcct cagtttcatc cccatcccca ggtcttccta
     2281 gacgctcagc acccccttct cccctgtgcc gtagtttgtc tcctgggact gggggaggag
     2341 tccgaggtgg ggttggttac ctgtcccgag gggaccctgt ccgggtcctt gctcggagag
     2401 tacggcctga tggctctgtg cagtacctgg ttgagtgggg aggagggggc atcttcggat
     2461 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     2521 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     2581 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2641 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2701 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2761 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2821 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2881 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2941 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3001 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3061 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3121 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3181 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3241 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3301 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3361 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3421 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3601 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3661 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3781 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3841 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4141 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4201 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4261 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4321 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4381 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4441 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4501 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4561 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4621 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4681 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4741 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4801 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4861 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4921 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4981 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5041 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5101 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5161 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5221 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5281 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5341 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5401 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5461 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5521 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5581 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5641 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5761 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5821 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5881 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5941 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6001 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6061 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6121 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6181 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6241 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6301 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6361 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6661 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6721 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6781 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6841 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6901 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6961 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7021 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7081 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7141 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7201 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7261 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7321 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7381 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7441 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7561 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7621 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7681 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7741 ggcttgaccg acaattgcat gaagaatctg cttagg
//
