LOCUS       sequence-194149-        8536 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 8536)
  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 8536)
  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 FGR
FEATURES             Location/Qualifiers
     source          1..8536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    868..1067
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1268..1684
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1614..1636
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1652..1668
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1702..1711
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1708..1749
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1774..1797
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1804..1828
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2399..2423)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2482..2943
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2538..2555)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2765..2784
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2948..3742
                     /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(3002..3021)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3612..3631
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3929..4398
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7165..7753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7654..7673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7907..7924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8028..8455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8138..8159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8350..8369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
       61 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      661 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
      721 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
      781 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
      841 gggaccaccg acccaccacc gggaggcaag ctggccagca acttatctgt gtctgtccga
      901 ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct
      961 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
     1021 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
     1081 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     1141 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     1201 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1261 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1321 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1381 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1441 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1501 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1561 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1621 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1681 cgccggccgg atccagaatt cgccaccatg gggtcttcaa aatctaaacc aaaggacccc
     1741 agccagcgcc ggcgcaggat ccgaggttac cttgactaca aagacgatga cgacaagcaa
     1801 ttgacaagtt tgtacaaaaa agttggcatg ggctgtgtgt tctgcaagaa attggagccg
     1861 gtggccacgg ccaaggagga tgctggcctg gaaggggact tcagaagcta cggggcagca
     1921 gaccactatg ggcctgaccc cactaaggcc cggcctgcat cctcatttgc ccacatcccc
     1981 aactacagca acttctcctc tcaggccatc aaccctggct tccttgatag tggcaccatc
     2041 aggggtgtgt cagggattgg ggtgaccctg ttcattgccc tgtatgacta tgaggctcga
     2101 actgaggatg acctcacctt caccaagggc gagaagttcc acatcctgaa caatactgaa
     2161 ggtgactggt gggaggctcg gtctctcagc tccggaaaaa ctggctgcat tcccagcaac
     2221 tacgtggccc ctgttgactc aatccaagct gaagagtggt actttggaaa gattgggaga
     2281 aaggatgcag agaggcagct gctttcacca ggcaaccccc agggggcctt tctcattcgg
     2341 gaaagcgaga ccaccaaagg tgcctactcc ctgtccatcc gggaccggga tcagacattg
     2401 ccaactttct tgtacaaagt ggtttgatct agtcgaccaa ttccggttat tttccaccat
     2461 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     2521 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     2581 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     2641 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     2701 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     2761 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     2821 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     2881 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2941 taataccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3001 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3061 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3121 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3181 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3241 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3301 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3361 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3421 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3481 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3541 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3601 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3661 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3721 ccttcttgac gagttcttct gagcgggact ctggggttcg catcgataaa ataaaagatt
     3781 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     3841 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     3901 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     3961 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     4021 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     4081 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     4141 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     4201 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     4261 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     4321 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     4381 gtcagcgggg gtctttcaca ggccagtggg gccgcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt gcggccgcga aaaggacaag
     5461 cagcgaaaat tcacgccccc ttgggaggtg gcggcatatg caaaggatag cactcccact
     5521 ctactactgg gtatcatatg ctgactgtat atgcatgagg atagcatatg ctacccggat
     5581 acagattagg atagcatata ctacccagat atagattagg atagcatatg ctacccagat
     5641 atagattagg atagcctatg ctacccagat ataaattagg atagcatata ctacccagat
     5701 atagattagg atagcatatg ctacccagat atagattagg atagcctatg ctacccagat
     5761 atagattagg atagcatatg ctatccagat atttgggtag tatatgctac ccatggcaac
     5821 attagcccac cgtgctctca gcgacctcgt gaatatgagg accaacaacc ctgtgcttgg
     5881 cgctcaggcg caagtgtgtg taatttgtcc tccagatcgc agcaatcgcg cccctatctt
     5941 ggcccgccca cctacttatg caggtattcc ccggggtgcc attagtggtt ttgtgggcaa
     6001 gtggtttgac cgcagtggtt agcggggtta caatcagcca agttattaca cccttatttt
     6061 acagtccaaa accgcagggc ggcgtgtggg ggctgacgcg tgcccccact ccacaatttc
     6121 aaaaaaaaga gtggccactt gtctttgttt atgggcccca ttggcgtgga gccccgttta
     6181 attttcgggg gtgttagaga caaccagtgg agtccgctgc tgtcggcgtc cactctcttt
     6241 ccccttgtta caaatagagt gtaacaacat ggttcacctg tcttggtccc tgcctgggac
     6301 acatcttaat aaccccagta tcatattgca ctaggattat gtgttgccca tagccataaa
     6361 ttcgtgtgag atggacatcc agtctttacg gcttgtcccc accccatgga tttctattgt
     6421 taaagatatt cagaatgttt cattcctaca ctagtattta ttgcccaagg ggtttgtgag
     6481 ggttatattg gtgtcatagc acaatgccac cactgaaccc cccgtccaaa ttttattctg
     6541 ggggcgtcac ctgaaacctt gttttcgagc acctcacata caccttactg ttcacaactc
     6601 agcagttatt ctattagcta aacgaaggag aatgaagaag caggcgaaga ttcaggagag
     6661 ttcactgccc gctccttgat cttcagccac tgcccttgtg actaaaatgg ttcactaccc
     6721 tcgtggaatc ctgaccccat gtaaataaaa ccgtgacagc tcatggggtg ggagatatcg
     6781 ctgttcctta ggaccctttt actaacccta attcgatagc atatgcttcc cgttgggtaa
     6841 catatgctat tgaattaggg ttagtctgga tagtatatac tactacccgg gaagcatatg
     6901 ctacccgttt agggttaaca agggggcctt ataaacacta ttgctaatgc cctcttgagg
     6961 gtccgcttat cggtagctac acaggcccct ctgattgacg ttggtgtagc ctcccgtagt
     7021 cttcctgggc ccctgggagg tacatgtcgg ccgcaaaagg atctaggtga agatcctttt
     7081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7921 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7981 ttaatgcaac tatggttggc gcgttgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     8041 tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa
     8101 atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact
     8161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     8221 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     8281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     8341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
     8401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgccgat
     8461 cttcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
     8521 tattggccat tgcata
//
