LOCUS       sequence-177675-        7273 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 7273)
  AUTHORS   Zhao JJ, Gjoerup OV, Subramanian RR, Cheng Y, Chen W, Roberts TM, 
            Hahn WC
  TITLE     Human mammary epithelial cell transformation through the activation 
            of phosphatidylinositol 3-kinase.
  JOURNAL   Cancer Cell. 2003 May . 3(5):483-95.
  PUBMED    12781366
REFERENCE   2  (bases 1 to 7273)
  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. 2003 May . 3(5):483-95."
COMMENT     Sequence Label: pBABE-puro SV40 LT
FEATURES             Location/Qualifiers
     source          1..7273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             12..481
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    546..745
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             946..1362
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1292..1314
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1330..1346
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1400..3526
                     /codon_start=1
                     /product="large T antigen from SV40 (simian virus 40)"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     primer_bind     complement(3607..3627)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3612..3941
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3792..3927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3951..4550
                     /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(3951..3970)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4447..4467
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4771..5240
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5370..5387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5541..6129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5621..5640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6289..7080,7081..7149))
                     /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     6912..6931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7150..7254)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgcag cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
       61 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
      121 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
      181 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
      241 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
      301 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg
      361 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
      421 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
      481 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
      541 gaggcaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
      601 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
      661 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
      721 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
      781 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
      841 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
      901 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
      961 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1021 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1081 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1141 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1201 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1261 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1321 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggccggat cccgcgaccg
     1381 gtcgaccagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     1441 ggaccttcta ggtcttgaaa ggagtgcctg ggggaatatt cctctgatga gaaaggcata
     1501 tttaaaaaaa tgcaaggagt ttcatcctga taaaggagga gatgaagaaa aaatgaagaa
     1561 aatgaatact ctgtacaaga aaatggaaga tggagtaaaa tatgctcatc aacctgactt
     1621 tggaggcttc tgggatgcaa ctgagattcc aacctatgga actgatgaat gggagcagtg
     1681 gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga
     1741 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     1801 ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag
     1861 aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa
     1921 aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat
     1981 actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa
     2041 attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag
     2101 tgccttgact agagatccat tttctgttat tgaggaaagt ttgccaggtg ggttaaagga
     2161 gcatgatttt aatccagaag aagcagagga aactaaacaa gtgtcctgga agcttgtaac
     2221 agagtatgca atggaaacaa aatgtgatga tgtgttgtta ttgcttggga tgtacttgga
     2281 atttcagtac agttttgaaa tgtgtttaaa atgtattaaa aaagaacagc ccagccacta
     2341 taagtaccat gaaaagcatt atgcaaatgc tgctatattt gctgacagca aaaaccaaaa
     2401 aaccatatgc caacaggctg ttgatactgt tttagctaaa aagcgggttg atagcctaca
     2461 attaactaga gaacaaatgt taacaaacag atttaatgat cttttggata ggatggatat
     2521 aatgtttggt tctacaggct ctgctgacat agaagaatgg atggctggag ttgcttggct
     2581 acactgtttg ttgcccaaaa tggattcagt ggtgtatgac tttttaaaat gcatggtgta
     2641 caacattcct aaaaaaagat actggctgtt taaaggacca attgatagtg gtaaaactac
     2701 attagcagct gctttgcttg aattatgtgg ggggaaagct ttaaatgtta atttgccctt
     2761 ggacaggctg aactttgagc taggagtagc tattgaccag tttttagtag tttttgagga
     2821 tgtaaagggc actggagggg agtccagaga tttgccttca ggtcagggaa ttaataacct
     2881 ggacaattta agggattatt tggatggcag tgttaaggta aacttagaaa agaaacacct
     2941 aaataaaaga actcaaatat ttccccctgg aatagtcacc atgaatgagt acagtgtgcc
     3001 taaaacactg caggccagat ttgtaaaaca aatagatttt aggcccaaag attatttaaa
     3061 gcattgcctg gaacgcagtg agtttttgtt agaaaagaga ataattcaaa gtggcattgc
     3121 tttgcttctt atgttaattt ggtacagacc tgtggctgag tttgctcaaa gtattcagag
     3181 cagaattgtg gagtggaaag agagattgga caaagagttt agtttgtcag tgtatcaaaa
     3241 aatgaagttt aatgtggcta tgggaattgg agttttagat tggctaagaa acagtgatga
     3301 tgatgatgaa gacagccagg aaaatgctga taaaaatgaa gatggtgggg agaagaacat
     3361 ggaagactca gggcatgaaa caggcattga ttcacagtcc caaggctcat ttcaggcccc
     3421 tcagtcctca cagtctgttc atgatcataa tcagccatac cacatttgta gaggttttac
     3481 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3541 ttgttgttaa cggggatccc agtgtggtgg tacgtaggaa ttcgccagca cagtggtcga
     3601 ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3661 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3781 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3841 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3901 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttacc atgaccgagt
     3961 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4021 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4081 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4141 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4201 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4261 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4321 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4381 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4441 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4501 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     4561 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgata aaataaaaga
     4621 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     4681 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     4741 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4801 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     4861 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     4921 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     4981 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     5041 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     5101 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     5161 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     5221 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     5281 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     5341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     6241 cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6301 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6361 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6421 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6481 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6541 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6601 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6661 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6721 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6781 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6841 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6901 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6961 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7021 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7081 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7141 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7201 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7261 tccccgaaaa gtg
//
