LOCUS       sequence-154447-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  AUTHORS   Wiedemeyer R, Brennan C, Heffernan TP, Xiao Y, Mahoney J, Protopopov
            A, Zheng H, Bignell G, Furnari F, Cavenee WK, Hahn WC, Ichimura K, 
            Collins VP, Chu GC, Stratton MR, Ligon KL, Futreal PA, Chin L
  TITLE     Feedback circuit among INK4 tumor suppressors constrains human 
            glioblastoma development.
  JOURNAL   Cancer Cell. 2008 Apr . 13(4):355-64.
  PUBMED    18394558
REFERENCE   2  (bases 1 to 5563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer 
            Cell. 2008 Apr . 13(4):355-64."
COMMENT     Sequence Label: pBabepuro3-p16Flag
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             11..480
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    545..744
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             945..1361
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1291..1313
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1329..1345
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1837..1860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1894..1914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1899..2228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2079..2214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2238..2837
                     /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(2238..2257)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2734..2754
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3060..3529
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3659..3676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3830..4418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4578..5369,5370..5438))
                     /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     5201..5220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5439..5543)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgcagc ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
       61 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
      121 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
      181 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
      241 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
      301 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
      361 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
      421 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
      481 tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg
      541 aggcaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt
      601 ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg
      661 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg
      721 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat
      781 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
      841 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt
      901 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca
      961 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     1021 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     1081 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     1141 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     1201 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
     1261 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
     1321 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggccggatc cgggctcgag
     1381 accatggagc cggcggctga ctggctggcc acggccgcgg cccggggtcg ggtagaggag
     1441 gtgcgggcgc tgctggaggc gggggcgctg cccaacgcac cgaatagtta cggtcggagg
     1501 ccgatccagg tcatgatgat gggcagcgcc cgagtggcgg agctgctgct gctccacggc
     1561 gcggagccca actgcgccga ccccgccact ctcacccgac ccgtgcacga cgctgcccgg
     1621 gagggcttcc tggacacgct ggtggtgctg caccgggccg gggcgcggct ggacgtgcgc
     1681 gatgcctggg gccgtctgcc cgtggacctg gctgaggagc tgggccatcg cgatgtcgca
     1741 cggtacctgc gcgcggctgc ggggggcacc agaggcagta accatgcccg catagatgcc
     1801 gcggaaggtc cctcagacat ccccgatgga tccatggact acaaagacga tgacgataaa
     1861 tagaagatct cccatcgatg gggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa
     1921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1981 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2041 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2101 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2161 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2221 tttgcaaaaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     2281 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     2341 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     2401 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     2461 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     2521 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     2581 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     2641 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     2701 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     2761 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     2821 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     2881 gaccccatgc atcgcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     2941 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3001 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     3061 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3121 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3181 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     3241 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3301 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3361 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     3421 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     3481 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     3541 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     3601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4501 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     4561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgc
//
