LOCUS       sequence-357942-        9274 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 9274)
  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 9274)
  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 PDK1
FEATURES             Location/Qualifiers
     source          1..9274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(430..1221,1222..1290))
                     /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     1053..1072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1291..1395)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1431..1900
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(2087..2881)
                     /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(2198..2217)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2808..2827
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_feature    2886..3347
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3045..3064)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3274..3291
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    3406..3430
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4739..4763)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4770..4793)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4818..4859)
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     regulatory      4856..4865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4883..5299)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4899..4915)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4931..4953)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5500..5699
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     promoter        complement(5920..6123)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5949..5969)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6124..6503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      6648..7075
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6734..6753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6944..6965
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7179..7196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7350..7938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7430..7449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc
       61 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
      121 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
      181 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
      241 taatctgtat ccgggtagca tatgctatcc tcatgcatat acagtcagca tatgataccc
      301 agtagtagag tgggagtgct atcctttgca tatgccgcca cctcccaagg gggcgtgaat
      361 tttcgctgct tgtccttttc gcggccgcaa atcaatctaa agtatatatg agtaaacttg
      421 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      481 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      541 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      601 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      661 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      721 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      781 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      841 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      901 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      961 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1021 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1081 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1141 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1201 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1261 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1321 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1381 aataggggtt ccgcgcacat ttccccgaaa agtgcggccc cactggcctg tgaaagaccc
     1441 ccgctgacgg gtagtcaatc actcagagga gaccctccca aggaacagcg agaccacaag
     1501 tcggatgcaa ctgcaagagg gtttattgga tacacgggta cccgggcgac tcagtcaatc
     1561 ggaggactgg cgccccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa
     1621 gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat
     1681 ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg ctgagggctg
     1741 gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca
     1801 gatatcctgt ttggcccata ttcagctgtt ccatctgttc ttggccctga gccggggcag
     1861 gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc tgttcctgac
     1921 cttgatctga acttctctat tctcagttat gtatttttcc atgccttgca aaatggcgtt
     1981 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
     2041 ctaaataaaa tcttttattt tatcgatgcg aaccccagag tcccgctcag aagaactcgt
     2101 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     2161 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     2221 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     2281 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     2341 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     2401 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     2461 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     2521 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     2581 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     2641 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     2701 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     2761 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     2821 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     2881 tggtattatc atcgtgtttt tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga
     2941 cgttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc agatcagatc
     3001 ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg cttgaggaga
     3061 gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt tgtgccgcct
     3121 ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg ccaggtgggg
     3181 ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa gcttccagag
     3241 gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac
     3301 ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag
     3361 acggcaatat ggtggaaaat aaccggaatt ggtcgactag atcaaaccac tttgtacaag
     3421 aaagttgggt aggcactgcg gaacgtcgtc atgtctttgg gttctctgct ggggacgcac
     3481 cagtcatcag cctcgtggtt ggtgttgtaa tgcttccagg cagctttgtt atacactggg
     3541 agtctttcta ttgagtctgt tgacagagcc ttaatgtaga taactgcatc tgtcccgtaa
     3601 ccctctaggg aatacagctt caggtctcct tggaagtatt gtgcgtaaag acgtgatatg
     3661 ggcaatccat aaccaaaacc agccagaggc actgcgcggg aggtctcaac acgaggtctt
     3721 ggtgcagttg aatacatgta gttgaaaagt ctgtcaattt tcctcaaagg aacgccacct
     3781 cctcggtcac tcatcttcac agtcaaatcc tcattaccca gcgtgacatg aacttgaata
     3841 ggggggtaaa cacctctgtt ggcatggtgt tccatagtgg ctctcattgc attcttgaaa
     3901 agttcaaaca ccatgtgata gagatgggat ggtacataaa ccacttgtat tggctgtcct
     3961 ggtgattttg catttagttc ttcaagttct agttcgggag agttaatata atacaaatca
     4021 cacagacgcc tagcattttc atagccatct ttaataactt caagtacatt gcagtttgga
     4081 tttatgcttc caatgtgttt tcgatgagat ggacttcctt tgccttttcc accaaacaat
     4141 aaagagtgct gattgagtaa cattctaatt gaaatgcgac tcatgtagaa tcgatccaaa
     4201 aagtactgaa cattctggct ggtgacagga tccaccccaa agctctcctt gtattcaatc
     4261 acaccctggg ccattgtggg aatgacatca ttgtgtcggt ttctgatccg tatcacagta
     4321 tctgtaaagt cataaatagc tttagcatcc tcagcacttt tgtccttaaa atcaagaagc
     4381 tcctgaagac tctggatata ccagctttgt accaattgaa cggatggtgt cctgagaaga
     4441 ttatctggaa ggagacttat ttctttcatt atatttgcca gtctgacagg caactcttgc
     4501 cgcagaaaca taaatgaggt cttttcacaa gcattcactg atccgaagtc caggaactgc
     4561 ttcatggaga gcggggacgg cgagaagcgc gcgtagaagt ccacctggcc cggaacgccg
     4621 cgctcggacg ccgggctgga gcccgagtcc gagctgaagc tgcggctgaa gccggcggcg
     4681 cgcagccccg ggcccgggcc ggccaaggcg gctccgcgaa gcagccgcgc cagcctcatg
     4741 ccaacttttt tgtacaaact tgtcaattgc ttgtcgtcat cgtctttgta gtcaaggtaa
     4801 cctcggatcc tgcgccggcg ctggctgggg tcctttggtt tagattttga agaccccatg
     4861 gtggcgaatt ctggatccgg ccggcgccta gagaaggagt gagggctgga taaagggagg
     4921 atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga cggggcggat ggaggaagag
     4981 gaggcggagg cttagggtgt acaaagggct tgacccaggg aggggggtca aaagccaagg
     5041 cttcccaggt cacgatgtag gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga
     5101 ccttgatctt aacctgggtg atgaggtctc ggttaaaggt gccgtctcgc ggccatccga
     5161 cgttaaaggt tggccattct gcagagcaga aggtaaccca acgtctcttc ttgacatcta
     5221 ccgactggtt gtgagcgatc cgctcgacat ctttccagtg acctaaggtc aaacttaagg
     5281 gagtggtaac agtctggccc taattttcag acaaatacag aaacacagtc agacagagac
     5341 aacacagaac gatgctgcag cgctgcagca gacaagacgc gcggcttcgg ttccaaaccg
     5401 aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt ctcgtctcct accagaacca
     5461 catatcctga cggggtcgga ttccacatcg actcccttcc tcaggtcggg ccacaaaaac
     5521 ggcccccaaa gtccctggga cgtctcccag ggttgcggcc gggtgttcag aactcgtcag
     5581 ttccaccacg ggtccgccag atacagagct agttagctaa ctagtaccga cgcaggcgca
     5641 taaaatcagt catagacact agacaatcgg acagacacag ataagttgct ggccagcttg
     5701 cctcccggtg gtgggtcggt ggtccctggg caggggtctc ccgatcccgg acgagccccc
     5761 aaatgaaaga cccccgctga cgggtagtca atcactcaga ggagaccctc ccaaggaaca
     5821 gcgagaccac aagtcggatg caactgcaag agggtttatt ggatacacgg gtacccgggc
     5881 gactcagtca atcggaggac tggcgcggtt cactaaacca gctctgctta tatagacctc
     5941 ccaccgtaca cgcctaccgc ccatttgcgt caacggggcg gggttattac gacattttgg
     6001 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     6061 tggaaatccc cgtgagtcaa accgctatcc acgcccattg gtgtactgcc aaaaccgcat
     6121 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccgtaa
     6181 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     6241 cggacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     6301 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     6361 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     6421 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     6481 ataactagtc aataatcaat gccaacatgg cggtcatatt ggacatgagc caatataaat
     6541 gtacatatta tgatatagat acaacgtatg caatggccaa tagccaatat tgatttatgc
     6601 tatataacca atgaataata tggctaatgg ccaatattga agatcggcgc gccctgtagc
     6661 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6721 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6781 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6841 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     6901 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6961 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     7021 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7081 aaaatattaa cgtttacaac gcgccaacca tagttgcatt aatgaatcgg ccaacgcgcg
     7141 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8041 atccttttgc ggccgacatg tacctcccag gggcccagga agactacggg aggctacacc
     8101 aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc
     8161 aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag
     8221 tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat
     8281 atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat
     8341 gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta
     8401 gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc
     8461 tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg
     8521 tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg
     8581 gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc
     8641 aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg
     8701 gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac
     8761 acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa
     8821 gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga
     8881 cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc
     8941 aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg
     9001 cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa
     9061 cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta
     9121 atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg
     9181 ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg
     9241 gtcctcatat tcacgaggtc gctgagagca cggt
//
