LOCUS       40924_505        3928 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3928)
  AUTHORS   Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.
  TITLE     Multiplexed and programmable regulation of gene networks with an 
            integrated RNA and CRISPR/Cas toolkit in human cells
  JOURNAL   Mol. Cell 54 (4), 698-710 (2014)
  PUBMED    24837679
REFERENCE   2  (bases 1 to 3928)
  AUTHORS   Perli SD.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2014) Electrical Engineering and Computer Science,
            Massachusetts Institute of Technology, 77, Massachusetts Avenue, 
            Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 3928)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3928)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
            date: "2014"; volume: "54"; issue: "4"; pages: "698-710"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-MAY-2014) Electrical Engineering and Computer Science, 
            Massachusetts Institute of Technology, 77, Massachusetts Avenue, 
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(620..724)
                     /label=AmpR promoter
     rep_origin      751..1206
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     enhancer        1395..1698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1699..1902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     ncRNA           1936..1972
                     /label=hammerhead ribozyme
     misc_RNA        1993..2068
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           2073..2140
                     /label=HDV ribozyme
     polyA_signal    2389..2436
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(2928..3516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3690..3928,1..619))
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
       61 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      361 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      721 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
      781 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      841 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      901 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      961 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1021 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1081 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1141 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     1201 caatttccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     1261 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     1321 tacttggagg tacctagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1381 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1441 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1501 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1561 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1621 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1681 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1741 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1801 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1861 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttatg aaccgtcaga
     1921 tccgagctcc gactactgat gagtccgtga ggacgaaacg agtaagctcg tctagtcgcg
     1981 tgtagcgaag cagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     2041 acttgaaaaa gtggcaccga gtcggtgctt ttggccggca tggtcccagc ctcctcgctg
     2101 gcgccggctg ggcaacatgc ttcggcatgg cgaatgggac cccgggatgc tagctaagcg
     2161 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     2221 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     2281 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     2341 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2401 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     2461 gctggcactc tgtcgatacc ccaccgagac gccattgggg ccaatacgcc cgcgtttctt
     2521 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     2581 gggcggcagg ccctgccata gcctcagctc gagtatgtcg accgatgccc ttgagagcct
     2641 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2701 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     2761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3121 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3901 cctgcaactt tatccgcctc catccagt
//
