LOCUS       sequence-461450-        9310 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 9310)
  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 9310)
  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 MAP2K5
FEATURES             Location/Qualifiers
     source          1..9310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    953..1152
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1353..1769
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1699..1721
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1737..1753
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1787..1796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1793..1834
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1859..1882
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1889..1913
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3258..3282)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3341..3802
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3397..3414)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3624..3643
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3807..4601
                     /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(3861..3880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4471..4490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4788..5257
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5293..5397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5398..5466,5467..6258)
                     /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(5616..5635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8024..8612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8513..8532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8766..8783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8887..9310,1..4))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8997..9018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9209..9228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcgccgatc ttcaatattg gccattagcc atattattca ttggttatat agcataaatc
       61 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
      121 ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta ttaatagtaa
      181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctggtttagt gaaccgcgcc agtcctccga ttgactgagt cgcccgggta
      781 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
      841 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
      901 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc
      961 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc
     1021 gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     1081 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     1141 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     1201 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     1261 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     1321 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     1381 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     1441 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1501 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1561 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1621 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1681 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1741 atccagccct cactccttct ctaggcgccg gccggatcca gaattcgcca ccatggggtc
     1801 ttcaaaatct aaaccaaagg accccagcca gcgccggcgc aggatccgag gttaccttga
     1861 ctacaaagac gatgacgaca agcaattgac aagtttgtac aaaaaagttg gcatgctgtg
     1921 gctagccctt ggcccctttc ctgccatgga gaaccaggtg ctggtaattc gcatcaagat
     1981 cccaaatagt ggcgcggtgg actggacagt gcactccggg ccgcagttac tcttcaggga
     2041 tgtgctggat gtgataggcc aggttctgcc tgaagcaaca actacagcat ttgaatatga
     2101 agatgaagat ggtgatcgaa ttacagtgag aagtgatgag gaaatgaagg caatgctgtc
     2161 atattattat tccacagtaa tggaacagca agtaaatgga cagttaatag agcctctgca
     2221 gatatttcca agagcctgca agcctcctgg ggaacggaac atacatggcc tgaaggtgaa
     2281 tactcgggcc ggaccctctc aacacagcag cccagcagtc tcagattcac ttccaagcaa
     2341 tagcttaaag aagtcttctg ctgaactgaa aaaaatacta gccaatggcc agatgaatga
     2401 acaagacata cgatatcggg acactcttgg tcatggcaac ggaggcacag tctacaaagc
     2461 atatcatgtc ccgagtggga aaatattagc tgtaaaggtc atactactag atattacact
     2521 ggaacttcag aagcaaatta tgtctgaatt ggaaattctt tataagtgcg attcatcata
     2581 tatcattgga ttttatggag cattttttgt agaaaacagg atttcaatat gtacagaatt
     2641 catggatggg ggatctttgg atgtatatag gaaaatgcca gaacatgtcc ttggaagaat
     2701 tgcagtagca gttgttaaag gccttactta tttgtggagt ttaaagattt tacatagaga
     2761 cgtgaagccc tccaatatgc tagtaaacac aagaggacag gttaagctgt gtgattttgg
     2821 agttagcact cagctggtga attctatagc caagacgtat gttggaacaa atgcttatat
     2881 ggcgcctgaa aggatttcag gggagcagta tggaattcat tctgatgtct ggagcttagg
     2941 aatctctttt atggagcttg ctcttgggag gtttccatat cctcagattc agaaaaacca
     3001 gggatcttta atgcctctcc agcttctgca gtgcattgtt gatgaggatt cgcccgtcct
     3061 tccagttgga gagttctcgg agccatttgt acatttcatc actcagtgta tgcgaaaaca
     3121 gccaaaagaa aggccagcac ctgaagaatt gatgggccac ccgttcatcg tgcagttcaa
     3181 tgatggaaat gccgccgtgg tgtccatgtg ggtgtgccgg gcgctggagg agaggcggag
     3241 ccagcagggg cccccgtgcc caactttctt gtacaaagtg gtttgatcta gtcgaccaat
     3301 tccggttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     3361 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     3421 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     3481 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     3541 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     3601 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     3661 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     3721 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     3781 cctttgaaaa acacgatgat aataccatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4621 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4681 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4741 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4801 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4861 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4921 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     4981 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     5041 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     5101 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     5161 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5221 tctgagtgat tgactacccg tcagcggggg tctttcacag gccagtgggg ccgcactttt
     5281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5401 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5461 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgatttg
     6301 cggccgcgaa aaggacaagc agcgaaaatt cacgccccct tgggaggtgg cggcatatgc
     6361 aaaggatagc actcccactc tactactggg tatcatatgc tgactgtata tgcatgagga
     6421 tagcatatgc tacccggata cagattagga tagcatatac tacccagata tagattagga
     6481 tagcatatgc tacccagata tagattagga tagcctatgc tacccagata taaattagga
     6541 tagcatatac tacccagata tagattagga tagcatatgc tacccagata tagattagga
     6601 tagcctatgc tacccagata tagattagga tagcatatgc tatccagata tttgggtagt
     6661 atatgctacc catggcaaca ttagcccacc gtgctctcag cgacctcgtg aatatgagga
     6721 ccaacaaccc tgtgcttggc gctcaggcgc aagtgtgtgt aatttgtcct ccagatcgca
     6781 gcaatcgcgc ccctatcttg gcccgcccac ctacttatgc aggtattccc cggggtgcca
     6841 ttagtggttt tgtgggcaag tggtttgacc gcagtggtta gcggggttac aatcagccaa
     6901 gttattacac ccttatttta cagtccaaaa ccgcagggcg gcgtgtgggg gctgacgcgt
     6961 gcccccactc cacaatttca aaaaaaagag tggccacttg tctttgttta tgggccccat
     7021 tggcgtggag ccccgtttaa ttttcggggg tgttagagac aaccagtgga gtccgctgct
     7081 gtcggcgtcc actctctttc cccttgttac aaatagagtg taacaacatg gttcacctgt
     7141 cttggtccct gcctgggaca catcttaata accccagtat catattgcac taggattatg
     7201 tgttgcccat agccataaat tcgtgtgaga tggacatcca gtctttacgg cttgtcccca
     7261 ccccatggat ttctattgtt aaagatattc agaatgtttc attcctacac tagtatttat
     7321 tgcccaaggg gtttgtgagg gttatattgg tgtcatagca caatgccacc actgaacccc
     7381 ccgtccaaat tttattctgg gggcgtcacc tgaaaccttg ttttcgagca cctcacatac
     7441 accttactgt tcacaactca gcagttattc tattagctaa acgaaggaga atgaagaagc
     7501 aggcgaagat tcaggagagt tcactgcccg ctccttgatc ttcagccact gcccttgtga
     7561 ctaaaatggt tcactaccct cgtggaatcc tgaccccatg taaataaaac cgtgacagct
     7621 catggggtgg gagatatcgc tgttccttag gaccctttta ctaaccctaa ttcgatagca
     7681 tatgcttccc gttgggtaac atatgctatt gaattagggt tagtctggat agtatatact
     7741 actacccggg aagcatatgc tacccgttta gggttaacaa gggggcctta taaacactat
     7801 tgctaatgcc ctcttgaggg tccgcttatc ggtagctaca caggcccctc tgattgacgt
     7861 tggtgtagcc tcccgtagtc ttcctgggcc cctgggaggt acatgtcggc cgcaaaagga
     7921 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7981 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8041 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8101 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8161 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8221 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8281 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8341 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8401 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8461 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8521 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8581 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8641 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8701 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8761 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     8821 ccgcgcgttg gccgattcat taatgcaact atggttggcg cgttgtaaac gttaatattt
     8881 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     8941 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     9001 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     9061 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     9121 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     9181 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     9241 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     9301 cgctacaggg
//
