LOCUS       sequence-381317-        6502 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 6502)
  AUTHORS   Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E, 
            Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y, 
            Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA, 
            Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA, 
            Meyerson M, Hahn WC
  TITLE     CDK8 is a colorectal cancer oncogene that regulates beta-catenin 
            activity.
  JOURNAL   Nature. 2008 Sep 25. 455(7212):547-51.
  PUBMED    18794900
REFERENCE   2  (bases 1 to 6502)
  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: "Nature. 
            2008 Sep 25. 455(7212):547-51."
COMMENT     Sequence Label: pBabe.puro.CDK8.flag
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             7..476
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    541..740
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             941..1357
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1287..1309
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1325..1341
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1368..1377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1377..1400
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2831..2851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2836..3165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3016..3151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3175..3774
                     /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(3175..3194)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3671..3691
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3995..4464
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4594..4611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5513..6304,6305..6373))
                     /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     6136..6155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6374..6478)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
       61 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
      121 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
      181 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
      241 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
      301 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
      361 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
      421 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
      481 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggc
      541 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
      601 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      661 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
      721 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
      781 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
      841 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
      901 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
      961 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     1021 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     1081 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     1141 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     1201 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     1261 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     1321 ctccctttat ccagccctca ctccttctct aggcgccggc cggatccgcc accatggact
     1381 acaaagacga tgacgacaag agatctatgg actatgactt taaagtgaag ctgagcagcg
     1441 agcgggagcg ggtcgaggac ctgtttgaat acgagggctg caaagttggc cgaggcactt
     1501 atggtcacgt ctacaaagcc aagaggaaag atgggaagga tgataaagac tatgctttaa
     1561 aacaaataga aggaactggg atctctatgt cggcatgtag agaaatagca ttacttcgag
     1621 agcttaagca tccaaacgtc atttctcttc aaaaggtgtt tctgtctcat gctgatagga
     1681 aggtgtggct tctgtttgac tatgctgaac atgacctctg gcatataatc aagtttcaca
     1741 gagcttctaa agcaaacaag aagccagttc agttacctcg gggaatggtg aagtcactat
     1801 tatatcagat cctagatggt attcactacc tgcatgctaa ctgggtgttg cacagagatt
     1861 tgaaacctgc taatatttta gttatgggtg aaggtcctga gcgaggaaga gtaaaaattg
     1921 ctgacatggg ctttgcccga ttatttaatt cacctttgaa gcctttagca gatttggatc
     1981 cagtggttgt tacattctgg taccgagccc ctgaactact tcttggagca aggcattata
     2041 ccaaagctat tgatatttgg gctatagggt gtatatttgc agaactacta acgtcagaac
     2101 caatatttca ctgtcgacaa gaggacatca aaactagtaa tccttatcac catgaccagc
     2161 tggacagaat attcaatgta atgggatttc ctgcagataa agattgggaa gatataaaaa
     2221 agatgcctga acattcaaca ttaatgaaag atttcagaag aaatacgtat accaactgca
     2281 gccttatcaa gtatatggaa aaacataaag ttaaaccaga tagtaaagca ttccacttgc
     2341 ttcagaagct gcttaccatg gacccaataa agcgaattac ctcagaacag gctatgcagg
     2401 acccctattt cttagaagac ccacttccta catcagacgt ttttgccggt tgtcaaatcc
     2461 cttacccaaa acgagaattt ttaacggaag aagaacctga tgacaaagga gacaaaaaga
     2521 accagcagca gcagcagggc aataaccaca ctaatggaac tggccaccca gggaatcaag
     2581 acagcagtca cacacaggga cccccgttga agaaagtgag agttgttcct cctaccacta
     2641 cctcaggtgg acttatcatg acctcagact atcagcgttc caatccacat gctgcctatc
     2701 ccaaccctgg accaagcaca tcacagccgc agagcagcat gggatactca gctacctccc
     2761 agcagcctcc acagtactca catcagacac atcggtactg agaattcgcc agcacagtgg
     2821 tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taccatgacc
     3181 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3241 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3301 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3361 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3421 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3481 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3541 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3601 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3661 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3721 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     3781 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa
     3841 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3901 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     3961 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     4021 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     4081 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4141 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4201 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4261 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4321 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     4381 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4441 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     4501 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     4561 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4621 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4681 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6481 catttccccg aaaagtgcca cc
//
