LOCUS       sequence-9516-ex        7783 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 7783)
  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 7783)
  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: PT2/C-fluc
FEATURES             Location/Qualifiers
     source          1..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..108
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     100..117
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     101..117
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     127..146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        127..145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        751..1130
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1133..1408
                     /label=chicken beta-actin promoter
     intron          1409..2417
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2341..2363
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2425..2444
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2472..4124
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2538..2556)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4273..4292)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4338..4393
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4392..4411)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(5096..5114)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5135..5151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5135..5151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5148..5170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5159..5175
                     /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(5183..5189,5190..5207,5208..5213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5228..5249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5366..5383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5537..6125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5617..5636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6296..7087,7088..7156))
                     /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     6919..6938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7157..7261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7288..7743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7375..7394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7585..7606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
       61 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
      121 cgcgcgtaat acgactcact atagggcgaa ttggagctcg gatccctata cagttgaagt
      181 cggaagttta catacactta agttggagtc attaaaactc gtttttcaac tactccacaa
      241 atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact ttgtgcatga
      301 cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata attcactgta
      361 tcacaattcc agtgggtcag aagtttacat acactaagtt gactgtgcct ttaaacagct
      421 tggaaaattc cagaaaatga tgtcatggct ttagaagcta gatctggcca tctagagcgg
      481 ccgcgcgcac tagtgaattc catggatatc aagctttttc cccgtatccc ccaggtgtct
      541 gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg
      601 tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag
      661 cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg
      721 ggggctttgg gggggggctg tccccgtgtc gacattgatt attgactagt tattaatagt
      781 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      841 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      901 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggactatt
      961 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1021 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1081 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ggtcgaggtg
     1141 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     1201 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggcgcgcgcc
     1261 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
     1321 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     1381 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg ttgccttcgc cccgtgcccc
     1441 gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     1501 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     1561 cgtttctttt ctgtggctgc gtgaaagcct taaagggctc cgggagggcc ctttgtgcgg
     1621 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggcc
     1681 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcgt
     1741 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggc tgcgagggga
     1801 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg
     1861 tcgggctgta acccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     1921 ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     1981 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     2041 cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     2101 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgg cggagccgaa
     2161 atctgggagg cgccgccgca ccccctctag cgggcgcggg cgaagcggtg cggcgccggc
     2221 aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccat
     2281 ctccagcctc ggggctgccg cagggggacg gctgccttcg ggggggacgg ggcagggcgg
     2341 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     2401 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     2461 gcaaagaatt catggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatccgc
     2521 tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc
     2581 ctggaacaat tgcttttaca gatgcacata tcgaggtgaa catcacgtac gcggaatact
     2641 tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca
     2701 gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat
     2761 ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca
     2821 gtatgaacat ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg caaaaaattt
     2881 tgaacgtgca aaaaaaatta ccaataatcc agaaaattat tatcatggat tctaaaacgg
     2941 attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta
     3001 atgaatacga ttttgtacca gagtcctttg atcgtgacaa aacaattgca ctgataatga
     3061 actcctctgg atctactggg ttacctaagg gtgtggccct tccgcataga actgcctgcg
     3121 tcagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga
     3181 ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga
     3241 tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ttacgatccc
     3301 ttcaggatta caaaattcaa agtgcgttgc tagtaccaac cctattttca ttcttcgcca
     3361 aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct tctgggggcg
     3421 cacctctttc gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt ccagggatac
     3481 gacaaggata tgggctcact gagactacat cagctattct gattacaccc gagggggatg
     3541 ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg
     3601 ataccgggaa aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga ggacctatga
     3661 ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat
     3721 ggctacattc tggagacata gcttactggg acgaagacga acacttcttc atagttgacc
     3781 gcttgaagtc tttaattaaa tacaaaggat accaggtggc ccccgctgaa ttggagtcga
     3841 tattgttaca acaccccaac atcttcgacg cgggcgtggc aggtcttccc gacgatgacg
     3901 ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
     3961 agatcgtgga ttacgtcgcc agtcaagtaa caaccgccaa aaagttgcgc ggaggagttg
     4021 tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag
     4081 agatcctcat aaaggccaag aagggcggaa agtccaaatt gtaaaatgta actgtattca
     4141 gcgatgacga aattcttagc tattgtaata ctctaggggg gatccactag aattcactcc
     4201 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4261 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4321 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4381 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4441 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     4501 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4561 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4621 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4681 actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct
     4741 tgtggaaggc tactcgaaat gtttgaccca agttaaacaa tttaaaggca atgctaccaa
     4801 atactaattg agtgtatgta aacttctgac ccactgggaa tgtgatgaaa gaaataaaag
     4861 ctgaaatgaa tcattctctc tactattatt ctgatatttc acattcttaa aataaagtgg
     4921 tgatcctaac tgacctaaga cagggaattt ttactaggat taaatgtcag gaattgtgaa
     4981 aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac ttccgacttc aactgtatag
     5041 ggatcctcta gctagagtcg acctcgaggg ggggcccggt acccagcttt tgttcccttt
     5101 agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7261 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     7321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7681 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     7741 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cga
//
