LOCUS       Exported                6249 bp ds-DNA     circular SYN 14-AUG-2017
DEFINITION  Cloning vector pCLIPf-H2B, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 12
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Sun L, Provost C, Ghosh I, Xu MQ.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-APR-2011) Research Department, New England Biolabs, 
            240 County Road, Ipswich, MA 01938, USA
REFERENCE   2  (bases 1 to 6249)
  AUTHORS   Addgene
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 14, 2017 from SnapGene 3.1.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6249
                     /organism="Cloning vector pCLIPf-H2B"
                     /mol_type="other DNA"
     promoter        251..818
                     /note="CMV immediate early promoter region"
     promoter        863..880
                     /note="T7 promoter (clockwise)"
     CDS             924..1940
                     /codon_start=1
                     /gene="H2B/CLIP10f"
                     /product="histone H2B / O-6-methylguanine-DNA 
                     methyltransferase fusion"
                     /note="H2B/CLIP10f"
                     /note="H2B sequence 924..1301, CLIP10f sequence 1356..1901;
                     CLIP10f optimized for mammalian expression; possible 
                     alternative start 774"
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAKASDIGAPAFKSVQTGEFTMDKDCEMKRTTLDSPLGKL
                     ELSGCEQGLHRIIFLGKGTSAADAVEVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEE
                     FPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISESHLAALVGNPAATAAVNTALDGN
                     PVPILIPCHRVVQGDSDVGPYLGGLAVKEWLLAHEGHRLGKPGLGPAGGSAFKLEVN"
     gene            924..1940
                     /gene="H2B/CLIP10f"
                     /note="H2B/CLIP10f"
     misc_RNA        2333..2915
                     /note="internal ribosome entry site (IRES) from 
                     encephalomyocarditis virus (ECMV); allows polycistronic 
                     expression of H2B/CLIP10f and neoR, and required for 
                     expression of neoR"
     CDS             2936..3739
                     /codon_start=1
                     /gene="aph(3')-II"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="aph(3')-II"
                     /note="neoR, kanR, nptII (confers resistance to neomycin 
                     and kanamycin)"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2936..3739
                     /gene="aph(3')-II"
                     /note="aph(3')-II"
     promoter        complement(4141..4170)
                     /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)
                     (counter-clockwise)"
     rep_origin      complement(4494..5082)
                     /direction=LEFT
                     /note="pUC19 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     CDS             complement(5253..6113)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5253..6113)
                     /gene="bla"
                     /note="bla"
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 gcttggtacc
      901 gagctcggat cgatatcgaa ttgatgccag agccagcgaa gtctgctccc gccccgaaaa
      961 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
     1021 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
     1081 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
     1141 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
     1201 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
     1261 ccgagggtac taaggccatc accaagtaca ccagcgctaa ggctagcgat atcggcgcgc
     1321 cagcatttaa atctgtacag accggtgaat tcaccatgga caaagactgc gaaatgaagc
     1381 gcaccaccct ggatagccct ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc
     1441 accgtatcat cttcctgggc aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc
     1501 cagccgccgt gctgggcgga ccagagccac tgatccaggc caccgcctgg ctcaacgcct
     1561 actttcacca gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt
     1621 tccagcagga gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg
     1681 gagaggtcat cagcgagagc cacctggccg ccctggtggg caatcccgcc gccaccgccg
     1741 ccgtgaacac cgccctggac ggaaatcccg tgcccattct gatcccctgc caccgggtgg
     1801 tgcagggcga cagcgacgtg gggccctacc tgggcgggct cgccgtgaaa gagtggctgc
     1861 tggcccacga gggccacaga ctgggcaagc ctgggctggg tcctgcaggc ggatccgcgt
     1921 ttaaactcga ggttaattaa tgagcggccg cttaagccgg ccgcatagat aactgatcca
     1981 gtgtgctgga attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc
     2041 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat
     2101 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac
     2161 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt
     2221 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag
     2281 gtcgagcatg catctagggc ggccaattcc gcccctctcc ccccccccct tttccctccc
     2341 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2401 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2461 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2521 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2581 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2641 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2701 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2761 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     2821 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     2881 tttgaaaaac acgatgataa gcttgccaca acccgggata attcctgcag ccaatatggg
     2941 atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3001 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3061 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3121 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3181 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3241 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3301 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3361 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3421 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3481 cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3541 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3601 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3661 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3721 tcttgacgag ttcttctgag gggatcaatt ctctagataa ctgatcataa tcagccatac
     3781 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3841 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3961 tggtttgtcc aaactcatca atgtatctta acgcgtcgag tgcattctag ttgtggtttg
     4021 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6241 cctgacgtc
//
