LOCUS       sequence-459969-        8611 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 8611)
  AUTHORS   Kang S, Dong S, Gu TL, Guo A, Cohen MS, Lonial S, Khoury HJ, Fabbro 
            D, Gilliland DG, Bergsagel PL, Taunton J, Polakiewicz RD, Chen J
  TITLE     FGFR3 Activates RSK2 to Mediate Hematopoietic Transformation through
            Tyrosine Phosphorylation of RSK2 and Activation of the MEK/ERK 
            Pathway.
  JOURNAL   Cancer Cell. 2007 Sep . 12(3):201-14.
  PUBMED    17785202
REFERENCE   2  (bases 1 to 8611)
  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 
            Cell. 2007 Sep . 12(3):201-14."
COMMENT     Sequence Label: pMSCV-puro-mycRSK2wt
FEATURES             Location/Qualifiers
     source          1..8611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             64..580
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    644..985
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1494..1518
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1542..1571
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(3808..3832)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3850..4349
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3885..3908)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4282..4301
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4370..4969
                     /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(4370..4389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4866..4886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5069..5583
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5745..5761
                     /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(5769..5775,5776..5793,5794..5799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5814..5835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5952..5969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6123..6711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6203..6222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6882..7673,7674..7742))
                     /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     7505..7524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7743..7847)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7915..7933
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7971..7993)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8093..8112
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8306..8328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8456..8475
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg
       61 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
      121 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
      181 cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      241 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
      301 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
      361 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
      421 aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtattcc
      481 caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt gggagggtct
      541 cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc accgagattt
      601 ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg ccagcggtcg
      661 tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattagatct ctcgaggtta tcaacaagtt
     1501 tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat ggaacaaaaa cttatttctg
     1561 aagaagatct gccgctggcg cagctggcgg acccgtggca gaagatggct gtggagagcc
     1621 cttccgacag cgcggagaat ggacagcaaa ttatggatga acctatggga gaggaggaga
     1681 ttaacccaca aactgaagaa ggcagtatca aagaaattgc aatcacacat catgtgaagg
     1741 aaggacatga aaaggcagat ccttcccagt ttgaactttt aaaagtatta gggcagggat
     1801 catttggaaa ggttttctta gttaaaaaaa tctcaggctc tgatgctaga cagctttatg
     1861 ccatgaaagt attaaagaag gccacgctga aagttcgaga ccgtgttcgg acaaaaatgg
     1921 aacgtgatat cttggtagaa gtcaatcacc ctttcattgt caaattgcat tacgcttttc
     1981 aaacggaagg gaagttgtat cttattttgg attttctcag gggcggagac ttgtttacac
     2041 gcttatccaa agaggtgatg ttcacagagg aagatgtcaa attctacttg gctgaacttg
     2101 cacttgcttt agaccatctt catagcctgg gaataatcta tagagactta aaaccagaaa
     2161 acatacttct tgatgaagaa ggtcacatca agttaactga ttttggctta agtaaggaat
     2221 ctattgatca tgagaagaag gcttattctt tttgtggcac tgtggaatac atggctccag
     2281 aagtagttaa ccgcagaggt cacactcaga gtgcggactg gtggtccttt ggtgtgttga
     2341 tgtttgaaat gctcactggt acactacctt tccaaggaaa agatcgtaaa gaaacaatga
     2401 ctatgattct taaagccaaa ctcgggatgc cacagtttct gagtcctgaa gcccagagtc
     2461 ttttacgaat gcttttcaaa cggaatcctg caaacagatt aggtgctgga ccagatggag
     2521 ttgaagaaat taaaagacat tcatttttct caacaataga ctggaataaa ctatatagaa
     2581 gagagattca cccacctttt aagcctgcaa ctggcagacc tgaagataca ttttattttg
     2641 atcctgagtt tactgcaaaa actcccaaag attcaccggg cattccacct agtgctaacg
     2701 cacatcagct ttttcggggg tttagttttg ttgctattac ctcagatgat gaaagccaag
     2761 ctatgcagac agttggtgtg cattcaattg ttcagcaatt acacagaaac agtattcagt
     2821 ttactgatgg atatgaagta aaagaggata ttggcgttgg ctcatactcc gtttgtaaga
     2881 gatgtataca taaagctaca aacatggagt ttgccgtgaa gattattgat aaaagcaaga
     2941 gagacccaac agaagagatt gaaattcttc ttcgctatgg acagcatcca aacatcatta
     3001 ccctaaagga tgtgtatgat gatggaaaat atgtgtatgt agtaacagaa cttatgaaag
     3061 gaggtgaatt gctggataag attcttagac agaagttttt ctcagagcga gaggccagcg
     3121 ctgtcctgtt tactataacc aaaactgttg agtatcttca tgcacaaggg gtggttcaca
     3181 gagacttgaa acctagcaac attctttatg tggatgagtc tggtaatcca gaatctattc
     3241 gaatttgtga ttttggcttt gcaaaacaac tgagagcaga aaatggtctt ctcatgactc
     3301 cttgttatac tgcaaatttt gttgcaccag aggttttaaa acggcaaggt tatgatgctg
     3361 cctgtgatat atggagtctt ggcgtcctcc tttatacaat gcttactggt tacactccat
     3421 ttgcaaatgg ccctgatgat actccagagg aaatactggc acgaataggt agtggaaaat
     3481 tctcactcag tggtggttac tggaattctg tttcagacac agcaaaggac ctggtgtcaa
     3541 agatgcttca tgtagatcct catcagagac tgacggctgc tctggtgctc agacatcctt
     3601 ggattgtcca ctgggaccaa ctaccacaat accaactaaa cagacaggat gcgccgcatc
     3661 tcgtaaaggg tgccatggca gctacgtact ctgctttaaa ccgcaatcag tccccagtct
     3721 tggaaccagt gggccgctcc actcttgctc agcggagagg gattaaaaaa atcacctcaa
     3781 cagccctgtg aaagggtggg cgcgccgacc cagctttctt gtacaaagtg gttgataacg
     3841 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     3901 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     3961 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     4021 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     4081 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     4141 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     4201 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     4261 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     4321 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta
     4381 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4441 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4501 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4561 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4621 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4681 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4741 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4801 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4861 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4921 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     4981 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat
     5041 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     5101 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     5161 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     5221 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     5281 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     5341 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     5401 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     5461 gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     5521 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     5581 catgggtaac agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta
     5641 agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata
     5701 gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta tccgctcaca
     5761 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5821 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5881 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5941 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6001 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6061 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6121 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6181 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6241 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6301 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6361 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6421 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6481 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6541 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6601 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6661 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6721 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6781 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6841 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6901 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6961 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7021 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7081 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7141 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7201 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7261 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7321 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7381 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7441 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7501 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7561 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7621 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7681 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7801 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7861 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     7921 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     7981 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     8041 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     8101 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     8161 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8221 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     8281 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
     8341 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     8401 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     8461 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     8521 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     8581 tagttttgac tcaacaatat caccagctga a
//
