LOCUS       Exported                6077 bp ds-DNA     circular SYN 14-AUG-2017
DEFINITION  Cloning vector pCLIPf-Cox8A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 8
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  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 6077)
  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..6077
                     /organism="Cloning vector pCLIPf-Cox8A"
                     /mol_type="other DNA"
     promoter        251..818
                     /note="CMV immediate early promoter region"
     promoter        863..880
                     /note="T7 promoter (clockwise)"
     CDS             929..1768
                     /codon_start=1
                     /gene="Cox8A/CLIP10f"
                     /product="human cytochrome c oxidase subunit 8A / 
                     O-6-methylguanine-DNA methyltransferase fusion"
                     /note="Cox8A/CLIP10f"
                     /note="Cox8A sequence 929..1135, CLIP10f sequence 
                     1184..1729"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLPPEGKLGIMELAVGLT
                     SCFVTFLLPAGWILSHLETYRRPEDIGAPAFKSVQTGEFTMDKDCEMKRTTLDSPLGKL
                     ELSGCEQGLHRIIFLGKGTSAADAVEVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEE
                     FPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISESHLAALVGNPAATAAVNTALDGN
                     PVPILIPCHRVVQGDSDVGPYLGGLAVKEWLLAHEGHRLGKPGLGPAGGSAFKLEVN"
     gene            929..1768
                     /gene="Cox8A/CLIP10f"
                     /note="Cox8A/CLIP10f"
     misc_RNA        2161..2743
                     /note="internal ribosome entry site (IRES) from 
                     encephalomyocarditis virus (ECMV); allows polycistronic 
                     expression of Cox8A/CLIP10f and neoR, and required for 
                     expression of neoR"
     CDS             2764..3567
                     /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            2764..3567
                     /gene="aph(3')-II"
                     /note="aph(3')-II"
     promoter        complement(3969..3998)
                     /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)
                     (counter-clockwise)"
     rep_origin      complement(4322..4910)
                     /direction=LEFT
                     /note="pUC19 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     CDS             complement(5081..5941)
                     /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(5081..5941)
                     /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 cgatggccgc agcttaccat gtccgtcctg acgccgctgc tgctgcgggg
      961 cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc aagatccatt cgttgccgcc
     1021 ggaggggaag cttgggatca tggaattggc cgttgggctt acctcctgct tcgtgacctt
     1081 cctcctgcca gcgggctgga tcctgtcaca cctggagacc tacaggaggc cagaggatat
     1141 cggcgcgcca gcatttaaat ctgtacagac cggtgaattc accatggaca aagactgcga
     1201 aatgaagcgc accaccctgg atagccctct gggcaagctg gaactgtctg ggtgcgaaca
     1261 gggcctgcac cgtatcatct tcctgggcaa aggaacatct gccgccgacg ccgtggaagt
     1321 gcctgcccca gccgccgtgc tgggcggacc agagccactg atccaggcca ccgcctggct
     1381 caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag ccctgcacca
     1441 cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc tgaaagtggt
     1501 gaagttcgga gaggtcatca gcgagagcca cctggccgcc ctggtgggca atcccgccgc
     1561 caccgccgcc gtgaacaccg ccctggacgg aaatcccgtg cccattctga tcccctgcca
     1621 ccgggtggtg cagggcgaca gcgacgtggg gccctacctg ggcgggctcg ccgtgaaaga
     1681 gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtc ctgcaggcgg
     1741 atccgcgttt aaactcgagg ttaattaatg agcggccgct taagccggcc gcatagataa
     1801 ctgatccagt gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac
     1861 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     1921 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca
     1981 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac
     2041 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc
     2101 aactgcaggt cgagcatgca tctagggcgg ccaattccgc ccctctcccc cccccccttt
     2161 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2221 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2281 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2341 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2401 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2461 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2521 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2581 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     2641 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2701 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggataat tcctgcagcc
     2761 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2821 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2881 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2941 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3001 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3061 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3121 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3181 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3241 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3301 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3361 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3421 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3481 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3541 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagataact gatcataatc
     3601 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3661 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3781 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt
     3841 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     3901 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6061 aagtgccacc tgacgtc
//
