LOCUS       sequence-222774-        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 largeTcDNA
FEATURES             Location/Qualifiers
     source          1..7273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             16..485
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    550..749
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             950..1366
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1296..1318
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1334..1350
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1404..3530
                     /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(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3616..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3955..4554
                     /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(3955..3974)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4451..4471
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4775..5244
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5374..5391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5625..5644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6293..7084,7085..7153))
                     /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     6916..6935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7154..7258)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
       61 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
      121 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
      181 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
      241 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
      301 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
      361 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
      421 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
      481 tttcatttgg gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca
      541 ccgggaggca agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac
      601 tgattttatg cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg
      661 tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt
      721 tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc
      781 aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
      841 tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt
      901 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
      961 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     1021 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     1081 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     1141 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     1201 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     1261 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     1321 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccgcg
     1381 accggtcgac cagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc
     1441 taatggacct tctaggtctt gaaaggagtg cctgggggaa tattcctctg atgagaaagg
     1501 catatttaaa aaaatgcaag gagtttcatc ctgataaagg aggagatgaa gaaaaaatga
     1561 agaaaatgaa tactctgtac aagaaaatgg aagatggagt aaaatatgct catcaacctg
     1621 actttggagg cttctgggat gcaactgaga ttccaaccta tggaactgat gaatgggagc
     1681 agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg
     1741 atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag
     1801 aagaccccaa ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta
     1861 atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca
     1921 agaaaattat ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata
     1981 acatactgtt ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc
     2041 aaaaattgtg tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt
     2101 atagtgcctt gactagagat ccattttctg ttattgagga aagtttgcca ggtgggttaa
     2161 aggagcatga ttttaatcca gaagaagcag aggaaactaa acaagtgtcc tggaagcttg
     2221 taacagagta tgcaatggaa acaaaatgtg atgatgtgtt gttattgctt gggatgtact
     2281 tggaatttca gtacagtttt gaaatgtgtt taaaatgtat taaaaaagaa cagcccagcc
     2341 actataagta ccatgaaaag cattatgcaa atgctgctat atttgctgac agcaaaaacc
     2401 aaaaaaccat atgccaacag gctgttgata ctgttttagc taaaaagcgg gttgatagcc
     2461 tacaattaac tagagaacaa atgttaacaa acagatttaa tgatcttttg gataggatgg
     2521 atataatgtt tggttctaca ggctctgctg acatagaaga atggatggct ggagttgctt
     2581 ggctacactg tttgttgccc aaaatggatt cagtggtgta tgacttttta aaatgcatgg
     2641 tgtacaacat tcctaaaaaa agatactggc tgtttaaagg accaattgat agtggtaaaa
     2701 ctacattagc agctgctttg cttgaattat gtggggggaa agctttaaat gttaatttgc
     2761 ccttggacag gctgaacttt gagctaggag tagctattga ccagttttta gtagtttttg
     2821 aggatgtaaa gggcactgga ggggagtcca gagatttgcc ttcaggtcag ggaattaata
     2881 acctggacaa tttaagggat tatttggatg gcagtgttaa ggtaaactta gaaaagaaac
     2941 acctaaataa aagaactcaa atatttcccc ctggaatagt caccatgaat gagtacagtg
     3001 tgcctaaaac actgcaggcc agatttgtaa aacaaataga ttttaggccc aaagattatt
     3061 taaagcattg cctggaacgc agtgagtttt tgttagaaaa gagaataatt caaagtggca
     3121 ttgctttgct tcttatgtta atttggtaca gacctgtggc tgagtttgct caaagtattc
     3181 agagcagaat tgtggagtgg aaagagagat tggacaaaga gtttagtttg tcagtgtatc
     3241 aaaaaatgaa gtttaatgtg gctatgggaa ttggagtttt agattggcta agaaacagtg
     3301 atgatgatga tgaagacagc caggaaaatg ctgataaaaa tgaagatggt ggggagaaga
     3361 acatggaaga ctcagggcat gaaacaggca ttgattcaca gtcccaaggc tcatttcagg
     3421 cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt
     3481 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3541 attgttgttg ttaacgggga tcccagtgtg gtggtacgta ggaattcgcc agcacagtgg
     3601 tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taccatgacc
     3961 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     4021 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     4081 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     4141 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     4201 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     4261 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     4321 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4381 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     4441 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4501 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     4561 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa
     4621 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     4681 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     4741 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     4801 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     4861 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4921 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4981 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     5041 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     5101 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     5161 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     5221 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     5281 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     5341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6601 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6901 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7261 catttccccg aaa
//
