LOCUS       40924_1519        8254 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector L071 RRI-Green, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8254)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Retroviral vectors for expression of shRNAs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8254)
  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 8254)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8254)
  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            "Unpublished"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..8254
                     /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..3708)
                     /label=EGFP
                     /note="enhanced GFP"
     promoter        complement(3734..4896)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        5105..5346
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    5355..5745
                     /note="stuffer fragment; lost during cloning of shRNAs
                     between unique BglII and AarI sites"
     misc_feature    5355
                     /label=U6 promoter-dependent transcription initiation
                     /note="U6 promoter-dependent transcription initiation"
     misc_feature    5802..5869
                     /label=R region
                     /note="R region"
     protein_bind    complement(6055..6071)
                     /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(6079..6109)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6124..6145)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6433..7021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7198..8055)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8056..8160)
                     /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 ccttgtacag
     3001 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     3061 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     3121 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     3181 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     3241 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     3301 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     3361 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     3421 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     3481 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     3541 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     3601 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     3661 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggatcgatc
     3721 ctaggccacg tgttcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag
     3781 gcttgagaat gaaccaagat ccaaactcaa aaaggccaaa ttccaaggag aattacatca
     3841 agtgccaagc tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat
     3901 aaaacccctc cccccaacct aaagacgacg tactccaaaa gctccagaac taatcgaggt
     3961 gcctggacgg cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg
     4021 cccctttcct ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt
     4081 ttgagccccc tggagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg
     4141 accgggccag ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca
     4201 gagcaggccg gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc
     4261 cccgcctcgc cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa
     4321 aaaccgaaat accagtgtgc agatcctggc ccgcatttac aagactatct tgccagaaaa
     4381 aaagcgtcgc agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga
     4441 gacaggcgcg aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct
     4501 caactcaagc acgaggcgaa ggggctccta aagcgcaagg cccgcccctg gctccagctc
     4561 gggatcaaga atcacgtact ggagccaggt ggaagtaatt caaggcacgc aagggccata
     4621 acccgtaaag aggccaggcc cgcgggaacc acacacggca cttacctgtg ttctggcggc
     4681 aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc
     4741 accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca
     4801 ccttctctag gcaccggttc aattgccgac ccctcccccc aacttctcgg ggactgtggg
     4861 cgatgtgcgc tctgcccact gacgggcacc ggagcctcac gctcgagagg ccgctcgacg
     4921 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     4981 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     5041 agaatagaaa agttcagatc aaggtcagga acagaggaac aggtcgacca aggtcgggca
     5101 ggaagagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     5161 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     5221 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     5281 tcatatgctt accgtaactt gaaagtactc tatcattgat agagttatat atcttgtgga
     5341 aaggacagat ctccgcatgc ggatccagga acaatggcag ctttcttgcc atcaaagctg
     5401 ctgatggcac atatattctt aatggtgact acactttgtc caccttagag caagacatta
     5461 tgtacaaagg tgttgtcttg aggtacagcg gctcctctgc ggcattggaa agaattcgca
     5521 gctttagccc tctcaaagag cccttgacca tccaggttct tactgtgggc aatgcccttc
     5581 gacctaaaat taaatacacc tacttcgtaa agaagaagaa ggaatctttc aatgctatcc
     5641 ccactttttc agcatgggtc attgaagagt ggggcgaatg ttctaagtca tgtgaattgg
     5701 gttggcagag aagactggta gaatgccgag acatcacctg catgcttttt ccatactttt
     5761 tttggcaagc ttacgcgtga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5821 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5881 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5941 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     6001 ggccatagta gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta
     6061 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6121 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6181 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6241 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6301 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6361 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6421 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6481 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6541 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6601 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6661 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6721 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6781 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6841 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6901 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6961 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7021 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7081 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7141 atgaagtttg cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7801 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8161 cacatttccc cgaaaagtgc caccagcttt gctcttagga gtttcctaat acatcccaaa
     8221 ctcaaatata taaagcattt gacttgttct atgc
//
