LOCUS       sequence-122724-        7464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Large T Antigen. Used to create 
            immortalized cell lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7464)
  AUTHORS   Hahn WC, Dessain SK, Brooks MW, King JE, Elenbaas B, Sabatini DM, 
            DeCaprio JA, Weinberg RA
  TITLE     Enumeration of the simian virus 40 early region elements necessary 
            for human cell transformation.
  JOURNAL   Mol Cell Biol 2002 Apr;22(7):2111-23.
  PUBMED    11884599
REFERENCE   2  (bases 1 to 7464)
  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: "Mol Cell 
            Biol"; date: "2002-04"; volume: "22(7)"; pages: "2111-23"
COMMENT     Sequence Label: pBABE-neo largeTcDNA
FEATURES             Location/Qualifiers
     source          1..7464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..475
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    540..739
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             940..1356
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1286..1308
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1324..1340
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1394..3520
                     /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(3601..3621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3606..3935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3786..3921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4292..5086
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4346..4365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4956..4975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5609..5628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5890..6359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(6474..7265,7266..7334))
                     /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     7097..7116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7335..7439)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
       61 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      121 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
      181 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
      241 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
      301 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
      361 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
      421 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
      481 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca
      541 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
      601 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
      661 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
      721 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
      781 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
      841 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
      901 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
      961 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1021 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1081 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1141 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1201 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1261 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1321 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccgcg accggtcgac
     1381 cagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     1441 tctaggtctt gaaaggagtg cctgggggaa tattcctctg atgagaaagg catatttaaa
     1501 aaaatgcaag gagtttcatc ctgataaagg aggagatgaa gaaaaaatga agaaaatgaa
     1561 tactctgtac aagaaaatgg aagatggagt aaaatatgct catcaacctg actttggagg
     1621 cttctgggat gcaactgaga ttccaaccta tggaactgat gaatgggagc agtggtggaa
     1681 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc
     1741 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa
     1801 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct
     1861 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat
     1921 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt
     1981 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg
     2041 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt
     2101 gactagagat ccattttctg ttattgagga aagtttgcca ggtgggttaa aggagcatga
     2161 ttttaatcca gaagaagcag aggaaactaa acaagtgtcc tggaagcttg taacagagta
     2221 tgcaatggaa acaaaatgtg atgatgtgtt gttattgctt gggatgtact tggaatttca
     2281 gtacagtttt gaaatgtgtt taaaatgtat taaaaaagaa cagcccagcc actataagta
     2341 ccatgaaaag cattatgcaa atgctgctat atttgctgac agcaaaaacc aaaaaaccat
     2401 atgccaacag gctgttgata ctgttttagc taaaaagcgg gttgatagcc tacaattaac
     2461 tagagaacaa atgttaacaa acagatttaa tgatcttttg gataggatgg atataatgtt
     2521 tggttctaca ggctctgctg acatagaaga atggatggct ggagttgctt ggctacactg
     2581 tttgttgccc aaaatggatt cagtggtgta tgacttttta aaatgcatgg tgtacaacat
     2641 tcctaaaaaa agatactggc tgtttaaagg accaattgat agtggtaaaa ctacattagc
     2701 agctgctttg cttgaattat gtggggggaa agctttaaat gttaatttgc ccttggacag
     2761 gctgaacttt gagctaggag tagctattga ccagttttta gtagtttttg aggatgtaaa
     2821 gggcactgga ggggagtcca gagatttgcc ttcaggtcag ggaattaata acctggacaa
     2881 tttaagggat tatttggatg gcagtgttaa ggtaaactta gaaaagaaac acctaaataa
     2941 aagaactcaa atatttcccc ctggaatagt caccatgaat gagtacagtg tgcctaaaac
     3001 actgcaggcc agatttgtaa aacaaataga ttttaggccc aaagattatt taaagcattg
     3061 cctggaacgc agtgagtttt tgttagaaaa gagaataatt caaagtggca ttgctttgct
     3121 tcttatgtta atttggtaca gacctgtggc tgagtttgct caaagtattc agagcagaat
     3181 tgtggagtgg aaagagagat tggacaaaga gtttagtttg tcagtgtatc aaaaaatgaa
     3241 gtttaatgtg gctatgggaa ttggagtttt agattggcta agaaacagtg atgatgatga
     3301 tgaagacagc caggaaaatg ctgataaaaa tgaagatggt ggggagaaga acatggaaga
     3361 ctcagggcat gaaacaggca ttgattcaca gtcccaaggc tcatttcagg cccctcagtc
     3421 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3481 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3541 ttaacgggga tcccagtgtg gtggtacgta ggaattcgcc agcacagtgg tcgaccctgt
     3601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3901 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tcacgctgcc gcaagcactc
     3961 agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg
     4021 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     4081 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     4141 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     4201 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     4261 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4321 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4381 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4441 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4501 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4561 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4621 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4681 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4741 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4801 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4861 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4921 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4981 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5041 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5101 ttcgcatcga tgcatcgatg cggcgggtgt ggtggttacg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcatcgataa aataaaagat tttatttagt ctccagaaaa
     5761 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     5821 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     5881 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     5941 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     6001 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     6061 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     6121 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     6181 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     6241 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     6301 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     6361 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     6421 tagtgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7441 acatttcccc gaaaagtgcc acct
//
