LOCUS       sequence-9517-ex        7148 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  AUTHORS   Wiesner SM, Decker SA, Larson JD, Ericson K, Forster C, Gallardo JL,
            Long C, Demorest ZL, Zamora EA, Low WC, SantaCruz K, Largaespada DA,
            Ohlfest JR
  TITLE     De novo induction of genetically engineered brain tumors in mice 
            using plasmid DNA.
  JOURNAL   Cancer Res. 2009 Jan 15. 69(2):431-9.
  PUBMED    19147555
REFERENCE   2  (bases 1 to 7148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
            2009 Jan 15. 69(2):431-9."
COMMENT     Sequence Label: pT/CMV-SV40-LgT
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..24)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13..237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(669..685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(669..685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(682..704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    693..709
                     /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(717..723,724..741,742..747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    762..783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(900..917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1071..1659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1151..1170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1830..2621,2622..2690))
                     /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     2453..2472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2691..2795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2863..2881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2919..2941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3041..3060
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3254..3276
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3268..3285
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3269..3285
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3684..4063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4064..4267
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4218..4238
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4313..4332
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4313..4331
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4481..4726,5073..6953)
                     /codon_start=1
                     /product="SV40 (simian virus 40) large T antigen"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     intron          4727..5072
                     /label=large T intron
                     /note="SV40 (simian virus 40) large T antigen intron"
     polyA_signal    6976..7110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7013..7032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7067..7086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
       61 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
      121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
      181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggaac
      241 cagcccatca caaagctctg acctcaatcc tatagaaagg aggaatgagc caaaattcac
      301 ccaacttatt gtgggaagct tgtggaaggc tactcgaaat gtttgaccca agttaaacaa
      361 tttaaaggca atgctaccaa atactaattg agtgtatgtt aacttctgac ccactgggaa
      421 tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc
      481 acattcttaa aataaagtgg tgatcctaac tgaccttaag acagggaatc tttactcgga
      541 ttaaatgtca ggaattgtga aaaagtgagt ttaaatgtat ttggctaagg tgtatgtaaa
      601 cttccgactt caactgtagg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
      661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1501 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2881 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2941 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3001 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     3061 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     3121 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3181 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     3241 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
     3301 acccgcagtt gaagtcggaa gtttacatac acttaagttg gagtcattaa aactcgtttt
     3361 tcaactacac cacaaatttc ttgttaacaa acaatagttt tggcaagtca gttaggacat
     3421 ctactttgtg catgacacaa gtcatttttc caacaattgt ttacagacag attatttcac
     3481 ttataattca ctgtatcaca attccagtgg gtcagaagtt tacatacact aagttgactg
     3541 tgcctttaaa cagcttggaa aattccagaa aatgatgtca tggctttaga agcttctgat
     3601 agactaattg acatcatttg agtcaattgg aggtgtacct gtggatgtat ttcaaggcga
     3661 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     3721 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     3781 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     3841 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggact atttacggta
     3901 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     3961 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4021 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     4081 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4141 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4201 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4261 cagagctctc tggctaacta gagaacccac tgcttaactg gcttatcgaa attaatacga
     4321 ctcactatag ggagacccaa gcttggtacc gagctcggat ccactagtaa cggccgccag
     4381 tgtgctggaa ttctgcagat atccatcaca ctggcggccg ccagtgtgat ggatatctgc
     4441 agaattcgcc ctctaggctt ttgcaaaaag ctttgcaaag atggataaag ttttaaacag
     4501 agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtgcct gggggaatat
     4561 tcctctgatg agaaaggcat atttaaaaaa atgcaaggag tttcatcctg ataaaggagg
     4621 agatgaagaa aaaatgaaga aaatgaatac tctgtacaag aaaatggaag atggagtaaa
     4681 atatgctcat caacctgact ttggaggctt ctgggatgca actgaggtat ttgcttcttc
     4741 cttaaatcct ggtgttgatg caatgtactg caaacaatgg cctgagtgtg caaagaaaat
     4801 gtctgctaac tgcatatgct tgctgtgctt actgaggatg aagcatgaaa atagaaaatt
     4861 atacaggaaa gatccacttg tgtgggttga ttgctactgc ttcgattgct ttagaatgtg
     4921 gtttggactt gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac
     4981 tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta
     5041 aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact gatgaatggg
     5101 agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa atgccatcta
     5161 gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg
     5221 tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat gctgtgttta
     5281 gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct gcactgctat
     5341 acaagaaaat tatggaaaaa tattctgtaa cctttataag taggcataac agttataatc
     5401 ataacatact gttttttctt actccacaca ggcatagagt gtctgctatt aataactatg
     5461 ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg ggttaataag gaatatttga
     5521 tgtatagtgc cttgactaga gatccatttt ctgttattga ggaaagtttg ccaggtgggt
     5581 taaaggagca tgattttaat ccagaagaag cagaggaaac taaacaagtg tcctggaagc
     5641 ttgtaacaga gtatgcaatg gaaacaaaat gtgatgatgt gttgttattg cttgggatgt
     5701 acttggaatt tcagtacagt tttgaaatgt gtttaaaatg tattaaaaaa gaacagccca
     5761 gccactataa gtaccatgaa aagcattatg caaatgctgc tatatttgct gacagcaaaa
     5821 accaaaaaac catatgccaa caggctgttg atactgtttt agctaaaaag cgggttgata
     5881 gcctacaatt aactagagaa caaatgttaa caaacagatt taatgatctt ttggatagga
     5941 tggatataat gtttggttct acaggctctg ctgacataga agaatggatg gctggagttg
     6001 cttggctaca ctgtttgttg cccaaaatgg attcagtggt gtatgacttt ttaaaatgca
     6061 tggtgtacaa cattcctaaa aaaagatact ggctgtttaa aggaccaatt gatagtggta
     6121 aaactacatt agcagctgct ttgcttgaat tatgtggggg gaaagcttta aatgttaatt
     6181 tgcccttgga caggctgaac tttgagctag gagtagctat tgaccagttt ttagtagttt
     6241 ttgaggatgt aaagggcact ggaggggagt ccagagattt gccttcaggt cagggaatta
     6301 ataacctgga caatttaagg gattatttgg atggcagtgt taaggtaaac ttagaaaaga
     6361 aacacctaaa taaaagaact caaatatttc cccctggaat agtcaccatg aatgagtaca
     6421 gtgtgcctaa aacactgcag gccagatttg taaaacaaat agattttagg cccaaagatt
     6481 atttaaagca ttgcctggaa cgcagtgagt ttttgttaga aaagagaata attcaaagtg
     6541 gcattgcttt gcttcttatg ttaatttggt acagacctgt ggctgagttt gctcaaagta
     6601 ttcagagcag aattgtggag tggaaagaga gattggacaa agagtttagt ttgtcagtgt
     6661 atcaaaaaat gaagtttaat gtggctatgg gaattggagt tttagattgg ctaagaaaca
     6721 gtgatgatga tgatgaagac agccaggaaa atgctgataa aaatgaagat ggtggggaga
     6781 agaacatgga agactcaggg catgaaacag gcattgattc acagtcccaa ggctcatttc
     6841 aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag
     6901 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     6961 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7021 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7081 ctcatcaatg tatcttatca tgtctggatc aagggcgaat tccagcacac tggcggccgt
     7141 tactagtg
//
