LOCUS       sequence-273879-        8860 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 8860)
  AUTHORS   Boehm JS, Zhao JJ, Yao J, Kim SY, Firestein R, Dunn IF, Sjostrom SK,
            Garraway LA, Weremowicz S, Richardson AL, Greulich H, Stewart CJ, 
            Mulvey LA, Shen RR, Ambrogio L, Hirozane-Kishikawa T, Hill DE, Vidal
            M, Meyerson M, Grenier JK, Hinkle G, Root DE, Roberts TM, Lander ES,
            Polyak K, Hahn WC
  TITLE     Integrative Genomic Approaches Identify IKBKE as a Breast Cancer 
            Oncogene (Kinase Library Clones)
  JOURNAL   Cell. 2007 Jun 15. 129(6):1065-1079.
  PUBMED    17574021
REFERENCE   2  (bases 1 to 8860)
  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: "Cell. 2007 
            Jun 15. 129(6):1065-1079."
COMMENT     Sequence Label: pWZL Neo Myr Flag CDK2
FEATURES             Location/Qualifiers
     source          1..8860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    805..1004
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1205..1621
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1551..1573
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1589..1605
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1639..1648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1645..1686
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1711..1734
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1741..1765
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2660..2684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2743..3204
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2799..2816)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3026..3045
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3209..4003
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3263..3282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3873..3892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4190..4659
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4695..4799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4800..4868,4869..5660)
                     /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     complement(5018..5037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7426..8014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7915..7934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8168..8185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8289..8716)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8399..8420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8611..8630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgcg
      601 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
      661 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
      721 ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg
      781 accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg
      841 tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg
      901 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
      961 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
     1021 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     1081 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
     1141 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
     1201 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1261 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1321 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1381 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1441 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1501 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1561 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1621 cggccggatc cagaattcgc caccatgggg tcttcaaaat ctaaaccaaa ggaccccagc
     1681 cagcgccggc gcaggatccg aggttacctt gactacaaag acgatgacga caagcaattg
     1741 acaagtttgt acaaaaaagt tggcatggag aacttccaaa aggtggaaaa gatcggagag
     1801 ggcacgtacg gagttgtgta caaagccaga aacaagttga cgggagaggt ggtggcgctt
     1861 aagaaaatcc gcctggacac tgagactgag ggtgtgccca gtactgccat ccgagagatc
     1921 tctctgctta aggagcttaa ccatcctaat attgtcaagc tgctggatgt catccacaca
     1981 gaaaataaac tctacctggt ttttgaattt ctgcaccaag atctcaagaa attcatggat
     2041 gcctctgctc tcactggcat tcctcttccc ctcatcaaga gctatctgtt ccagctgctc
     2101 cagggcctag ctttctgcca ttctcatcgg gtcctccacc gagaccttaa acctcagaat
     2161 ctgcttatta acacagaggg ggccatcaag ctagcagact ttggactagc cagagctttt
     2221 ggagtccctg ttcgtactta cacccatgag gtggtgaccc tgtggtaccg agctcctgaa
     2281 atcctcctgg gctgcaaata ttattccaca gctgtggaca tctggagcct gggctgcatc
     2341 tttgctgaga tggtgactcg ccgggcccta ttccctggag attctgagat tgaccagctc
     2401 ttccggatct ttcggactct ggggacccca gatgaggtgg tgtggccagg agttacttct
     2461 atgcctgatt gcaagccaag tttccccaag tgggcccggc aagattttag taaagttgta
     2521 cctcccctgg atgaagatgg acggagcttg ttatcgcaaa tgctgcacta cgaccctaac
     2581 aagcggattt cggccaaggc agccctggct caccctttct tccaggatgt gaccaagcca
     2641 gtaccccatc ttcgactctg cccaactttc ttgtacaaag tggtttgatc tagtcgacca
     2701 attccggtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2761 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     2821 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     2881 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     2941 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3001 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     3061 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3121 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     3181 ttcctttgaa aaacacgatg ataataccat gattgaacaa gatggattgc acgcaggttc
     3241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4021 gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     4081 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     4141 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     4201 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     4261 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4321 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     4381 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4441 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4501 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     4561 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4621 cctctgagtg attgactacc cgtcagcggg ggtctttcac aggccagtgg ggccgcactt
     4681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 tgcggccgcg aaaaggacaa gcagcgaaaa ttcacgcccc cttgggaggt ggcggcatat
     5761 gcaaaggata gcactcccac tctactactg ggtatcatat gctgactgta tatgcatgag
     5821 gatagcatat gctacccgga tacagattag gatagcatat actacccaga tatagattag
     5881 gatagcatat gctacccaga tatagattag gatagcctat gctacccaga tataaattag
     5941 gatagcatat actacccaga tatagattag gatagcatat gctacccaga tatagattag
     6001 gatagcctat gctacccaga tatagattag gatagcatat gctatccaga tatttgggta
     6061 gtatatgcta cccatggcaa cattagccca ccgtgctctc agcgacctcg tgaatatgag
     6121 gaccaacaac cctgtgcttg gcgctcaggc gcaagtgtgt gtaatttgtc ctccagatcg
     6181 cagcaatcgc gcccctatct tggcccgccc acctacttat gcaggtattc cccggggtgc
     6241 cattagtggt tttgtgggca agtggtttga ccgcagtggt tagcggggtt acaatcagcc
     6301 aagttattac acccttattt tacagtccaa aaccgcaggg cggcgtgtgg gggctgacgc
     6361 gtgcccccac tccacaattt caaaaaaaag agtggccact tgtctttgtt tatgggcccc
     6421 attggcgtgg agccccgttt aattttcggg ggtgttagag acaaccagtg gagtccgctg
     6481 ctgtcggcgt ccactctctt tccccttgtt acaaatagag tgtaacaaca tggttcacct
     6541 gtcttggtcc ctgcctggga cacatcttaa taaccccagt atcatattgc actaggatta
     6601 tgtgttgccc atagccataa attcgtgtga gatggacatc cagtctttac ggcttgtccc
     6661 caccccatgg atttctattg ttaaagatat tcagaatgtt tcattcctac actagtattt
     6721 attgcccaag gggtttgtga gggttatatt ggtgtcatag cacaatgcca ccactgaacc
     6781 ccccgtccaa attttattct gggggcgtca cctgaaacct tgttttcgag cacctcacat
     6841 acaccttact gttcacaact cagcagttat tctattagct aaacgaagga gaatgaagaa
     6901 gcaggcgaag attcaggaga gttcactgcc cgctccttga tcttcagcca ctgcccttgt
     6961 gactaaaatg gttcactacc ctcgtggaat cctgacccca tgtaaataaa accgtgacag
     7021 ctcatggggt gggagatatc gctgttcctt aggacccttt tactaaccct aattcgatag
     7081 catatgcttc ccgttgggta acatatgcta ttgaattagg gttagtctgg atagtatata
     7141 ctactacccg ggaagcatat gctacccgtt tagggttaac aagggggcct tataaacact
     7201 attgctaatg ccctcttgag ggtccgctta tcggtagcta cacaggcccc tctgattgac
     7261 gttggtgtag cctcccgtag tcttcctggg cccctgggag gtacatgtcg gccgcaaaag
     7321 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7381 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7441 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7501 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7561 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7621 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7681 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7741 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7801 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7861 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7921 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7981 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8041 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8101 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     8161 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     8221 ccccgcgcgt tggccgattc attaatgcaa ctatggttgg cgcgttgtaa acgttaatat
     8281 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     8341 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     8401 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8461 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     8521 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8581 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     8641 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8701 gccgctacag ggcgcgccga tcttcaatat tggccattag ccatattatt cattggttat
     8761 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     8821 tacatttata ttggctcatg tccaatatga ccgccatgtt
//
