LOCUS       40924_1524        8215 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector L087 RRI-Red, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8215)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Retroviral vectors for expression of shRNAs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8215)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2008) Biomedicine, University of Bergen, Jonas 
            Lies vei 91, Bergen 5009, Norway
REFERENCE   3  (bases 1 to 8215)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8215)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JAN-2008) Biomedicine, University of Bergen, Jonas Lies vei 91, 
            Bergen 5009, Norway"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             571..746
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     CDS             1240..1656
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1666..2044
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     polyA_signal    complement(2063..2270)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(2901..2985)
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(2992..3666)
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     promoter        complement(3695..4857)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        5066..5307
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    5316..5706
                     /note="stuffer fragment; lost during cloning of shRNAs
                     between unique BglII and AarI sites"
     misc_feature    5316
                     /label=U6 promoter-dependent transcription initiation
                     /note="U6 promoter-dependent transcription initiation"
     misc_feature    5763..5830
                     /label=R region
                     /note="R region"
     protein_bind    complement(6016..6032)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6040..6070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6085..6106)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6394..6982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7159..8016)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8017..8121)
                     /label=AmpR promoter
ORIGIN
        1 cctagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcat
      541 gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc cctcactcgg
      601 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
      661 gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg
      721 actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc
      781 agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct
      841 ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag
      901 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttcgga acacccggcc
      961 gcaaccctgg gagacgtccc agggacttcg ggggccgttt ttgtggcccg acctgagtcc
     1021 aaaaatcccg atcgttttgg actctttggt gcacccccct tagaggaggg atatgtggtt
     1081 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     1141 tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
     1201 ctgactgtgt ttctgtattt gtctgaaaat atgggcccgg gccagactgt taccactccc
     1261 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1321 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1381 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1441 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1501 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1561 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1621 tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata tgagatcgat
     1681 cttatatggg gcacccccgc cccttgtaaa cttccctgac cctgacatga caagagttac
     1741 taacagcccc tctctccaag ctcacttaca ggctctctac ttagtccagc acgaagtctg
     1801 gagacctctg gcggcagcct accaagaaca actggaccga ccggtggtac ctcaccctta
     1861 ccgagtcggc gacacagtgt gggtccgccg acaccagact aagaacctag aacctcgctg
     1921 gaaaggacct tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag acggcatcgc
     1981 agcttggata cacgccgccc acgtgaaggc tgccgacccc gggggtggac catcctctag
     2041 actgccggat ctcgaaggat cttccccacc atgcctgcta ttgtcttccc aatcctcccc
     2101 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     2161 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     2221 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggtcta
     2281 gcggccgctc tagatcaggc accgggcttg cgggtcatgc accaggtcgc gcggtccttc
     2341 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     2401 gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
     2461 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     2521 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     2581 cgcggccagc cgggaaccgc tcaactcggc catgcgcgga ccgatctcgg cgaacaccgc
     2641 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     2701 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     2761 ccgctcgatg tggcggtccg ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     2821 ggcggcgagg gtgcgtacgg cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
     2881 cttgtactct gtcattgatg tggtgggccc agggttggac tcgacgtctc ccgcaagttt
     2941 aagaaggtca aaattcaaca gctgaccgcc accgctgcca cctccgaatt cggcgccggt
     3001 ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg gtgtagtcct cgttgtggga
     3061 ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc agctgcacgg gcttcttggc
     3121 catgtaggtg gtcttgacct cggcgtcgta gtggccgccg tccttcagct tcagcctcat
     3181 cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggtgg aggcctccca
     3241 gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt
     3301 caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc
     3361 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa
     3421 gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtact ggaactgagg
     3481 ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt
     3541 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     3601 cacggagccc tccatgcgca ccttgaagcg catgaactcc ttgatgacgt cctcggagga
     3661 ggccaccatg gtggatcgat cctaggccac gtgttcacga cacctgaaat ggaagaaaaa
     3721 aactttgaac cactgtctga ggcttgagaa tgaaccaaga tccaaactca aaaaggccaa
     3781 attccaagga gaattacatc aagtgccaag ctggcctaac ttcagtctcc acccactcag
     3841 tgtggggaaa ctccatcgca taaaacccct ccccccaacc taaagacgac gtactccaaa
     3901 agctccagaa ctaatcgagg tgcctggacg gcgcccggta ctccgtggag tcacatgaag
     3961 cgacggctga ggacggaaag gcccctttcc tttgtgtggg tgactcaccc gcccgctctc
     4021 ccgagcgccg cgtcctccat tttgagcccc ctggagcagg gccgggaagc ggccatcttt
     4081 ccgctcacgc aactggtgcc gaccgggcca gccttgccgc ccagggcggg gcgatacacg
     4141 gcggcgcgag gccaggcacc agagcaggcc ggccagcttg agactacccc cgtccgattc
     4201 tcggtggccg cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc
     4261 cgtcgccgcc cgcggcccca aaaaccgaaa taccagtgtg cagatcctgg cccgcattta
     4321 caagactatc ttgccagaaa aaaagcgtcg cagcaggtca tcaaaaattt taaatggcta
     4381 gagacttatc gaaagcagcg agacaggcgc gaaggtgcca ccagattcgc acgcggcggc
     4441 cccagcgccc aggccaggcc tcaactcaag cacgaggcga aggggctcct aaagcgcaag
     4501 gcccgcccct ggctccagct cgggatcaag aatcacgtac tggagccagg tggaagtaat
     4561 tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc
     4621 acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
     4681 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
     4741 ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga cccctccccc
     4801 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagcctca
     4861 cgctcgagag gccgctcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     4921 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4981 gcatggaaaa atacataact gagaatagaa aagttcagat caaggtcagg aacagaggaa
     5041 caggtcgacc aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca
     5101 tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat
     5161 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     5221 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtact ctatcattga
     5281 tagagttata tatcttgtgg aaaggacaga tctccgcatg cggatccagg aacaatggca
     5341 gctttcttgc catcaaagct gctgatggca catatattct taatggtgac tacactttgt
     5401 ccaccttaga gcaagacatt atgtacaaag gtgttgtctt gaggtacagc ggctcctctg
     5461 cggcattgga aagaattcgc agctttagcc ctctcaaaga gcccttgacc atccaggttc
     5521 ttactgtggg caatgccctt cgacctaaaa ttaaatacac ctacttcgta aagaagaaga
     5581 aggaatcttt caatgctatc cccacttttt cagcatgggt cattgaagag tggggcgaat
     5641 gttctaagtc atgtgaattg ggttggcaga gaagactggt agaatgccga gacatcacct
     5701 gcatgctttt tccatacttt ttttggcaag cttacgcgtg agcccacaac ccctcactcg
     5761 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     5821 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     5881 actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt
     5941 tcttaagtat ttacattaaa tggccatagt agttcattat ggacagcgca gaaagagctg
     6001 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6061 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7081 ctagatcctt ttaaattaaa aatgaagttt gcgcaaatca atctaaagta tatatgagta
     7141 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7321 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7681 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7741 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7981 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8041 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8101 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccaccagctt tgctcttagg
     8161 agtttcctaa tacatcccaa actcaaatat ataaagcatt tgacttgttc tatgc
//
