LOCUS       sequence-9948-ex        7509 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  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 7509)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009 
            Aug 21. 138(4):673-84."
COMMENT     Sequence Label: pMSCV Cyclin D2-short
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2652..3151
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2687..2710)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3084..3103
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3172..3771
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3172..3191)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3668..3688
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3871..4385
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4554..4570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4554..4570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4567..4589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4578..4594
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(4602..4608,4609..4626,4627..4632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4647..4668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4785..4802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4956..5544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5036..5055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5715..6506,6507..6575))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6338..6357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6576..6680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6748..6766
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6804..6826)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6926..6945
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7139..7161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7289..7308
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7508..7509,1..515)
                     /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 gatcttattt agccaaagga aggaggtcag
     1441 gggaacgctc tcccctcccc ttccaaaaaa caaaaacaga aaaacccttt tccaggccgg
     1501 ggaaagcagg agggagaggg gccgccgggc tggccatgga gctgctgtgc cacgaggtgg
     1561 acccggtccg cagggccgtg cgggaccgca acctgctccg agacgaccgc gtcctgcaga
     1621 acctgctcac catcgaggag cgctaccttc cgcagtgctc ctacttcaag tgcgtgcaga
     1681 aggacatcca accctacatg cgcagaatgg tggccacctg gatgctggag gtctgtgagg
     1741 aacagaagtg cgaagaagag gtcttccctc tggccatgaa ttacctggac cgtttcttgg
     1801 ctggggtccc gactccgaag tcccatctgc aactcctggg tgctgtctgc atgttcctgg
     1861 cctccaaact caaagagacc agcccgctga ccgcggagaa gctgtgcatt tacaccgaca
     1921 actccatcaa gcctcaggag ctgctggagt gggaactggt ggtgctgggg aagttgaagt
     1981 ggaacctggc agctgtcact cctcatgact tcattgagca catcttgcgc aagctgcccc
     2041 agcagcggga gaagctgtct ctgatccgca agcatgctca gaccttcatt gctctgtgtg
     2101 ccaccgactt taagtttgcc atgtacccac cgtcgatgat cgcaactgga agtgtgggag
     2161 cagccatctg tgggctccag caggatgagg aagtgagctc gctcacttgt gatgccctga
     2221 ctgagctgct ggctaagatc accaacacag acgtggattg tctcaaagct tgccaggagc
     2281 agattgaggc ggtgctcctc aatagcctgc agcagtaccg tcaggaccaa cgtgacggat
     2341 ccaagtcgga ggatgaactg gaccaagcca gcacccctac agacgtgcgg gatatcgacc
     2401 tgtgaggatg ccagttgggc cgaaagagag agacgcgtcc ataatctggt ctcttcttct
     2461 ttctggttgt ttttgttctt tgtgttttag ggtgaaactt aaaaaaaaaa ttctgccccc
     2521 acctagatca tatttaaaga tcttttagaa gtgagagaaa aaggtcctac gaaaacggaa
     2581 taataaaaag catttggtgc ctatttgaag tacagcataa gggaatcccc tcgaggttaa
     2641 cgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     2701 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     2761 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     2821 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     2881 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     2941 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     3001 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     3061 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     3121 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag
     3181 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3241 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3301 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3361 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3421 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3481 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3541 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3601 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3661 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3721 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     3781 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat aaaataaaag
     3841 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     3901 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     3961 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     4021 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc
     4081 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     4141 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     4201 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     4261 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     4321 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     4381 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat
     4441 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa
     4501 tagttcatta tggacagcgc agaagagctg gggagaatta attcgtaatc atggtcatag
     4561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5701 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5761 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5821 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5881 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5941 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6001 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6061 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6121 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6181 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6241 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6301 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6361 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6421 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6481 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6541 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6721 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6781 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6841 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6901 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6961 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     7021 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     7081 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     7141 cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca
     7201 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     7261 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     7321 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat
     7381 ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     7441 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     7501 aggggggaa
//
