LOCUS       pMSCV16URP-U6-sh        7602 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Lentiviral vector derived from murine embryonic stem cell virus, for
            barcoded shRNA libraries, with TagRFP and puromycin resistance 
            markers.
ACCESSION   .
VERSION     .
KEYWORDS    pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..515
                     /label=MSCV
                     /note="Murine embryonic stem cell virus promoter including
                     the enhancer and promoter region of PCMV virus and 5' 
                     untranslated region of dl-587rev retrovirus"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     promoter        1438..1678
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    1684..1713
                     /label=cloning site
                     /note="cloning site"
                     /note="shRNA cloning site"
     promoter        1872..2271
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2284..2994
                     /label=TagRFP
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
     CDS             3001..3054
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=2A peptide from Thosea asigna virus capsid
                     protein
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3061..3657
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             3736..4161
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     promoter        4427..4756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(5049..5637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5811..6668)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6669..6773)
                     /label=AmpR promoter
     LTR             7601..7602
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga cggccgcccc cttcaccgag
     1441 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     1501 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     1561 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     1621 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     1681 aacaccgacg tcttcttttt tgaagacact tcggactgta gaactctgaa cctcgagcaa
     1741 taaaagaatg gtacagtgca gaaacaatag tagacataat agcaacagac atacaaacta
     1801 aagaatctgc gttgttgtcg gtgctcgttc tctgctcttc acgctactga attcatcacc
     1861 ggttcttcga aggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac
     1921 ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt ccgcccggac
     1981 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2041 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2101 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2161 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2221 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tctagacgcc
     2281 accatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
     2341 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
     2401 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     2461 gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc
     2521 atccccgact tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca
     2581 tacgaggacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     2641 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
     2701 aaaacactcg gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa
     2761 ggcagaagcg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag
     2821 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
     2881 gaccacagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag
     2941 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacaaact taatggatcc
     3001 gagggcagag gaagccttct aacatgcggt gacgtggagg agaatcccgg cccttccggg
     3061 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     3121 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     3181 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     3241 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     3301 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     3361 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     3421 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     3481 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     3541 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     3601 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     3661 atctaggtcg acgcctatag agtacgagcc atagataaaa taaaagattt tatttagtct
     3721 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3781 gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc
     3841 aggaacagat ggaacaggct cgaccctaga gaaccatcag atgtttccag ggtgccccaa
     3901 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3961 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     4021 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     4081 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     4141 cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga
     4201 ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg atgacggtga
     4261 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4321 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4381 gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt gtcagttagg
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4561 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4621 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4681 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4741 cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca gattgtactg
     4801 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4861 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4921 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4981 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6121 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6421 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6661 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6721 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6781 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6841 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6901 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6961 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     7021 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     7081 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     7141 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     7201 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     7261 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     7321 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     7381 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     7441 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     7501 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     7561 taaaataaaa gattttattt agtctccaga aaaagggggg aa
//
