LOCUS       sequence-394648-        8334 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 8334)
  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 8334)
  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 Res.
            2009 Jan 15. 69(2):431-9."
COMMENT     Sequence Label: PT3.5/CMV-EGFRvIII
FEATURES             Location/Qualifiers
     source          1..8334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(99..118)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    164..219
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(218..237)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(916..936)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(916..934)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(955..971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(955..971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(968..990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    979..995
                     /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(1003..1009,1010..1027,1028..1033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1048..1069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1186..1203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1357..1945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1437..1456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2116..2907,2908..2976))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2977..3081)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3108..3563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3195..3214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3405..3426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3689..3711
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3703..3720
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3704..3720
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3730..3749
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3730..3748
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        4171..4474
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4475..4678
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4629..4649
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5069..5485
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5415..5437
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5453..5469
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 tcgagctcaa gcttcgaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
       61 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
      121 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
      181 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
      241 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
      301 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
      361 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
      421 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
      481 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
      541 agatccctcg acctgcagcc caagcttgtg gaaggctact cgaaatgttt gacccaagtt
      601 aaacaattta aaggcaatgc taccaaatac taattgagtg tatgtaaact tctgacccac
      661 tgggaatgtg atgaaagaaa taaaagctga aatgaatcat tctctctact attattctga
      721 tatttcacat tcttaaaata aagtggtgat cctaactgac ctaagacagg gaatttttac
      781 taggattaaa tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc taaggtgtat
      841 gtaaacttcc gacttcaact gtatagggat cctctagcta gagtcgaggg ggggcccggt
      901 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
      961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1501 taccggatac ctgtccgcct ttctcccttc gggaaccgtg gcgctttctc atagctcacg
     1561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1861 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1921 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1981 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2161 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2221 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2281 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2341 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2401 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2461 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2521 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaacttggc
     2581 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2641 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2701 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2761 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2821 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2881 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2941 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3001 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3061 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     3121 cggcgcatta agcgcggcgg gagtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3181 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3301 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3421 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3481 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3541 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     3601 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     3661 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     3721 gagcgcgcgt aatacgactc actatagggc gaattggagc tcggtacccg ggggcgcgcc
     3781 ggatccatat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
     3841 cgtttttcaa ccactccaca aatttcttgt taacaaacta tagttttggc aagtcggtta
     3901 ggacatctac tttgtgcatg acacaagtaa tttttccaac aattgtttac agacagatta
     3961 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
     4021 tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc tttagaagct
     4081 tgcatgcctg caggtttaaa cagtcgactc tagttattaa tagtaatcaa ttacggggtc
     4141 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     4201 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     4261 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     4321 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     4381 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     4441 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     4501 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     4561 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     4621 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     4681 tttagtgaac cgtcagatcc gctagcgcta ccggttctag aacctaggga gctctggtac
     4741 ccactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
     4801 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
     4861 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
     4921 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
     4981 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
     5041 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     5101 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     5161 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     5221 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     5281 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     5341 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     5401 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     5461 agccctcact ccttctctag gcgccggaat tcgttggccg catgcgaccc tccgggacgg
     5521 ccggggcagc gctcctggcg ctgctggctg cgctctgccc ggcgagtcgg gctctggagg
     5581 aaaagaaagg taattatgtg gtgacagatc acggctcgtg cgtccgagcc tgtggggccg
     5641 acagctatga gatggaggaa gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc
     5701 gcaaagtgtg taacggaata ggtattggtg aatttaaaga ctcactctcc ataaatgcta
     5761 cgaatattaa acacttcaaa aactgcacct ccatcagtgg cgatctccac atcctgccgg
     5821 tggcatttag gggtgactcc ttcacacata ctcctcctct ggatccacag gaactggata
     5881 ttctgaaaac cgtaaaggaa atcacagggt ttttgctgat tcaggcttgg cctgaaaaca
     5941 ggacggacct ccatgccttt gagaacctag aaatcatacg cggcaggacc aagcaacatg
     6001 gtcagttttc tcttgcagtc gtcagcctga acataacatc cttgggatta cgctccctca
     6061 aggagataag tgatggagat gtgataattt caggaaacaa aaatttgtgc tatgcaaata
     6121 caataaactg gaaaaaactg tttgggacct ccggtcagaa aaccaaaatt ataagcaaca
     6181 gaggtgaaaa cagctgcaag gccacaggcc aggtctgcca tgccttgtgc tcccccgagg
     6241 gctgctgggg cccggagccc agggactgcg tctcttgccg gaatgtcagc cgaggcaggg
     6301 aatgcgtgga caagtgcaac cttctggagg gtgagccaag ggagtttgtg gagaactctg
     6361 agtgcataca gtgccaccca gagtgcctgc ctcaggccat gaacatcacc tgcacaggac
     6421 ggggaccaga caactgtatc cagtgtgccc actacattga cggcccccac tgcgtcaaga
     6481 cctgcccggc aggagtcatg ggagaaaaca acaccctggt ctggaagtac gcagacgccg
     6541 gccatgtgtg ccacctgtgc catccaaact gcacctacgg atgcactggg ccaggtcttg
     6601 aaggctgtcc aacgaatggg cctaagatcc cgtccatcgc cactgggatg gtgggggccc
     6661 tcctcttgct gctggtggtg gccctgggga tcggcctctt catgcgaagg cgccacatcg
     6721 ttcggaagcg cacgctgcgg aggctgctgc aggagaggga gcttgtgggg cctcttacac
     6781 ccagtggaga agctcccaac caagctctct tgaggatctt gaaggaaact gaattcaaaa
     6841 agatcaaagt gctgggctcc ggtgcgttcg gcacggtgta taagggactc tggatcccag
     6901 aaggtgagaa agttaaaatt cccgtcgcta tcaaggaatt aagagaagca acatctccga
     6961 aagccaacaa ggaaatcctc gatgaagcct acgtgatggc cagcgtggac aacccccacg
     7021 tgtgccgcct gctgggcatc tgcctcacct ccaccgtgca gctcatcacg cagctcatgc
     7081 ccttcggctg cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc
     7141 tgctcaactg gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg
     7201 tgcaccgtga cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca
     7261 cagattttgg gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag
     7321 gcaaagtgcc tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc
     7381 agagtgatgt ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc
     7441 catatgacgg aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc
     7501 ctcagccacc catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag
     7561 acgcagatag tcgcccaaag ttccgtgagt tgatcatcga attctccaaa atggcccgag
     7621 acccccagcg ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag
     7681 actccaactt ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg
     7741 acgagtacct catcccacag cagggcttct tcagcagccc ctccacgtca cggactcccc
     7801 tcctgagctc tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg
     7861 ggctgcaaag ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca
     7921 caggcgcctt gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa
     7981 accagtccgt tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc
     8041 ctctgaaccc cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg
     8101 gcaaccccga gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc
     8161 ctgcccactg ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc
     8221 aggacttctt tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa
     8281 atgcagaata cctaagggtc gcgccacaaa gcagtgaatt tattggagca tgac
//
