LOCUS       40924_32380        7627 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Retroviral backbone for cloning and gene expression. Select with 
            puromycin or GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7627)
  AUTHORS   Mayr C, Bartel DP
  TITLE     Widespread shortening of 3'UTRs by alternative cleavage and 
            polyadenylation activates oncogenes in cancer cells.
  JOURNAL   Cell. 2009 Aug 21. 138(4):673-84.
  PUBMED    19703394
REFERENCE   2  (bases 1 to 7627)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7627)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009 
            Aug 21. 138(4):673-84."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(676..692)
                     /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(700..730)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(745..766)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(883..900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1054..1642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1816..2673)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2674..2778)
                     /label=AmpR promoter
     primer_bind     2846..2864
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(2902..2924)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3024..3043
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     3237..3259
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3387..3406
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     LTR             3606..4122
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     misc_feature    4186..4527
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             4594..5010
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     promoter        5045..5544
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5565..6161
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6229..6805
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             6806..7522
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             7627
                     /label=3' LTR
                     /note="3' 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 gtatccaata
      421 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
      481 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
      541 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
      601 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
      661 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
      901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     2821 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3001 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3061 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     3121 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3181 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3241 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
     3301 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     3361 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     3421 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
     3481 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     3541 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     3601 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     3661 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     3721 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3781 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     3841 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3901 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     3961 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtatt
     4021 cccaataaag cctcttgctg tttgcatccg aatcgtggac tcgctgatcc ttgggagggt
     4081 ctcctcagat tgattgactg cccacctcgg gggtctttca tttggaggtt ccaccgagat
     4141 ttggagaccc ctgcccaggg accaccgacc cccccgccgg gaggtaagct ggccagcggt
     4201 cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg tttgcgcctg
     4261 cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     4321 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
     4381 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
     4441 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
     4501 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
     4561 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     4621 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     4681 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     4741 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     4801 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     4861 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     4921 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     4981 tatccagccc tcactccttc tctaggcgcc ggaattagat ctctcgaggt taacgaattc
     5041 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     5101 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     5161 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     5221 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     5281 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     5341 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     5401 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     5461 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     5521 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     5581 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     5641 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     5701 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     5761 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     5821 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     5881 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5941 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     6001 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     6061 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     6121 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     6181 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc caattccgcc ccccccccct
     6241 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     6301 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     6361 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     6421 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     6481 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     6541 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     6601 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     6661 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     6721 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     6781 acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt
     6841 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg
     6901 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     6961 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     7021 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     7081 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     7141 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     7201 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     7261 tatcatggcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat
     7321 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     7381 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     7441 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     7501 cggcatggac gagctgtaca agtaatgaat taattaagaa ttatcaagct tatcgatacc
     7561 gtcgacctgc agccaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag
     7621 gggggaa
//
